CN102548406A

CN102548406A - Process for the synthesis of 4,5,6,7-tetrachloro-3',6'-dihydroxy-2',4',5'7'-tetraiodo-3h-spiro[isobenzofuran-1,9'-xanthen]-3-one (rose bengal) and related xanthenes

Info

Publication number: CN102548406A
Application number: CN2010800415067A
Authority: CN
Inventors: J·辛格; E·A·沃奇特; T·C·斯科特; M·卢兹; K·巴比艾克
Original assignee: Provectus Biopharmaceuticals Inc
Current assignee: General vectus pharmaceutical technology company
Priority date: 2009-09-18
Filing date: 2010-09-17
Publication date: 2012-07-04
Anticipated expiration: 2030-09-17
Also published as: EP2464225A4; US9422260B2; MX2012003063A; HK1172212A1; US20130274322A1; ES2548575T3; JP5671040B2; HK1172501A1; JP2013505261A; US8530675B2; EP2464225A1; EP2464225B1; CN102548406B; CA2771988A1; EP2464225B8; US20110071217A1; KR20120091088A; WO2011035161A1; KR101494055B1; CA2771988C

Abstract

A new process for the manufacture of iodinated xanthenes in high purity includes a cyclization step followed by an iodination step. No extraction, chromatographic or solvent concentration steps are required, and the intermediate as well as final compounds are isolated via filtration or similar means. The process requires a single organic solvent, and the steps are completed at temperatures below 100 DEG C. The exclusion of chloride ions, of chloride free-radicals, hypochlorite ions, or hypochlorous acid as reagents or from reagents that may generate these species in situ in the presence of oxidants, prevents undesirable impurity formation. Several new compounds have been conceived and isolated using these methods. These new compounds are also formed into new medicaments.

Description

Synthesize 4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 ', the method for 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone (rose-red) and relevant xanthene

Cross reference with related application

The application requires in the rights and interests of the provisional application sequence number 61/243,701 of submission on September 18th, 2009.

Invention field

The present invention relates to prepare or method synthetic and rose-red, rose-red lactone of transport disengaging height purifying and relevant xanthene, and relate to highly purified rose-red, rose-red lactone and relevant xanthene.The synthetic method that relates in one aspect to preparation iodate fluorescein derivative of the present invention, said derivative contains 2% weight at most, preferably less than the independent organic impurities of 0.15% weight.In 0.15% or 1500ppm level or lower relevant with medicinal effectiveness, reason is the qualified threshold that International Conference of Harmonisation (ICH) guide is deferred in its representative with Control of Impurities.Therefore, another aspect of the present invention relates to the medicinal effectiveness and the evaluation of noval chemical compound disclosed herein, specially designs above-mentioned synthesizing for this purpose and controls its formation.

Background of invention

Fluorescein structural motif and it is believed that and at first be described in Baeyer (Berichte.1871:4,555) in 1871 from a step cyclisation of phthalic anhydride and resorcinol.Graebe (Annalen.1887:18,318) it is believed that it is to use the halogenation phthalic anhydride to do the cyclisation substrate first, mentions in its report and uses the excessive acid anhydrides (1.3 equivalent) of relative resorcinol.The iodate of dichlorofluorescein appeared in the document in 1887, and it is reported in Le Royer (Annalen.1887:238,359).In 20th century, several uses of fluorescein analog appear.This compounds has been used as the structural unit of textile dye, biology dyestuff, non-volatile memory unit, thermal imaging substrate and food color and cosmetics colouring agent.For example, the erythrosine (D&C Nos.11 and 12) of erythrosine (FD&C No.3) and part iodate is as food, medicine and cosmetic dyes.A kind of special tetraiodo xanthene, rose-red, it is visual to be used for eye disease, and is used as liver function medical diagnosis agent with the radio-labeled form, appears in the American Pharmacopeia in nineteen sixty-five.

Yet, the cyclisation of constructing rosy xanthene core since 1880 technology (high temperature melting in the opening still) not essence improve, be perhaps still very extensively with modifying the fluorescein primitive even so to synthetic non-halogenated analog.Known synthetic method produces the impurity of a series of uncertain and inferior signs, and it comprises residual solvent, inorganic compound and organic compound derived from reaction or degradation process.These impurity of many historical purposes in commercial Application, food dye or diagnostics allow.For example, the Code of Federal Regulations (CFR) of the U.S. allows the impurity level of FD&C No.3 (erythrosine) to be no more than 1% monoiod(in)ate impurity and is no more than the fluorescein of other lower iodate of 9%.The residual impurity that CFR also allows cyclisation step to produce; Such as the phthalic acid of part iodate and resorcinol (for example referring to Kamikura, Shokuhin Eiseigaku Zasshi 1985:26,243 with people such as Wada; Food.Add.Contam.2004:21,1137).

Above-mentioned historical colouring agent specification is different from the requirement of modern International Conference of Harmonisation (ICH) guide for novel drugs fully; It requires report 0.05% or higher impurity; Comprehensive identification level 0.1% or higher any organic impurities; And, and the restriction to inorganic impurity is provided further greater than the thorough toxicology specification of any impurity of 0.15%, particularly to the strictness restriction of residual solvent.So, will this compounds introduced under the situation of health with therapeutic dose, what become more and more important is good, the predictable and reproducible synthetic method of control.During synthesis step or in purifying, produce plurality of impurities is in the afore mentioned rules unpredictablely, and particularly for potential parenteral drug product preparaton, institute is unallowed.

In order to prepare the rose-red of reagent grade, American Pharmacopeia XXII recommends to come purifying rose-red with HCl via acid/alkaline treatment.The inventor finds very surprisingly; Under the situation that exists oxidant such as hydrogen peroxide or potassium hydrogen peroxymonosulfate; Cause side reaction with containing the agent treated iodate fluorescein that maybe can produce moisture cl anion, wherein one or more in the I substituting group can be Cl by trans halogenation.This also can betide and exist under chloride free radical, hypochlorite ion or the hypochlorous situation.This side reaction during preparation is rose-red does not have advance report, therefore must carefully control cyclisation step, iodate step and any purification schemes with this undesirable side reaction of prevention.

Describe in advance and the usually upper wording " halogen " of mentioning though iodate fluorescein analog is existing, not having any to seem during these are formerly open can be directly from the synthetic substituted fluorescein of iodo-xanthene of iodo-resorcinol.In addition, there is not any to seem in formerly disclosing and in molecule, has at least one iodine, and do not have any to require the medicinal application that must rely on iodine of these compounds of protection.Because fluorescein is used for non-therapeutic purposes in a large number; Almost do not have and mention: describe and summarize the needed method of high-purity activity drug ingedient of this compounds of enforcement preparation, and identify, characterize and the synthetic method that can serve as the secondary by-products of human therapeutic agent or colouring agent about information of the present invention.The iodate xanthene as embodiment be included in upperly various open in, and rose-red be to be described in 1880 the compound of knowing first.Yet these lists of references do not have separating or evaluation of any auxiliary product of describing the trans halogenation that is made up of at least one I on the xanthene core and at least one Cl substituting group, perhaps disclose or point out the possibility of its existence.The inventor has found that these products can be present in the rose-red commercial sample with as many as 2% weight.In addition; Formerly list of references than the xanthene pollutant that hangs down iodate (is for example inferred; In present CFR 74.303, allow to be no more than 9% for the dyestuff of erythrosine proof), but these lists of references do not have any proposition can be at 2 ' or 4 ' with the structure of the substituted triiodide form of hydrogen or its atropisomer or title (Fig. 1 q and the 1r that vide infra, and referring to the U.S. patent No. 6; The discussion of isomery is changeed in the resistance of 649,769 pairs of these skeletons).Any instruction is not formerly arranged in the list of references or enlighten said separation or make and to synthesize independent I/Cl substituting group around the xanthene core.The present invention identifies, characterizes and establish and controls the synthetic to satisfy the method that is used for the needed standard of medicinal application of these accessory substances effectively.In addition, the inventive method avoid these accessory substances undesirable formation, avoid need using multiple solvent and strict each reagent of control, these all improve processing, yield, purity and the feasibility of the method that is used for medicinal usage.

Brief summary of the invention

The present invention relates to control the material and the method for impurity level of iodate xanthene that preparation is used for formula 3 and the formula 4 of medicinal usage, relate to the method that prepare these iodate xanthene compounds, and disclose relevant but the trans halogenation impurity of expectation in advance not.The present invention also relates to be used to prepare their material and method, especially prepare method with the erythrosine (FD&C 3) that is suitable for medicinal purity, the rose-red iodate xanthene relevant with other.In one embodiment, this method is only used single organic solvent, and low temperature (less than 100 ℃) and requirement use very limited additive and conditioning agent to avoid forming impurity.Avoiding forming trans halogenation impurity can for example realize like this: in reactant mixture, particularly the cl anion in the reactant mixture during the iodate step is limited to less than 1500ppm.Do not need extraction, chromatogram or solvent concentration step, and intermediate and final compound can be via filtering or similar means be able to separate.The present invention also relates to specific analog and new compound, particularly can via preceding text disclosed method preparation or avoid those.These compounds have been able to separate and confirm, and are applicable to that medicine, medical science, cosmetics and colouring agent use.

In another embodiment, the method for preparing formula 3 compounds is provided,

The R of each position wherein ₁Be C independently ₁-C ₄Alkyl, halogen or H; And R ₂, R ₃, R ₄And R ₅Be I, F, Cl, C independently ₁-C ₄Alkyl or H, wherein R ₂, R ₃, R ₄Or R ₅In at least one is I, and the R of each position ₆Be H or C independently ₁-C ₄Alkyl; The aqueous solution existence that is included in alkali is following to formula 3 compounds, wherein the R of each position ₁Be C independently ₁-C ₄Alkyl, halogen or H; R ₂, R ₃, R ₄And R ₅Be F, Cl, C independently ₁-C ₄Alkyl or H and R ₂, R ₃, R ₄Or R ₅In at least one is H, with iodine (I ₂) reaction, to replace R with I ₂, R ₃, R ₄Or R ₅In at least one.

Another embodiment relates to the method for preparing formula 3 compounds, wherein the R of each position ₁Be C independently ₁-C ₄Alkyl, halogen or H; R ₂, R ₃, R ₄And R ₅Be halogen independently, C ₁-C ₄Alkyl or H; And the R of each position ₆Be H or C independently ₁-C ₄Alkyl; Comprise formula 1 compound,

The R of each position wherein ₁Be halogen, C independently ₁-C ₄Alkyl or H react with formula 2 compounds

R wherein ₇, R ₈, R ₁₀Be F, Cl, C independently ₁-C ₄Alkyl or H; And R ₉Be H.

Another embodiment relates to the method for preparing formula 3 derivatives, wherein the R of each position ₁Be halogen or H independently; R ₂, R ₃, R ₄And R ₅Be I, Br, Cl, F or H, wherein R independently ₂, R ₃, R ₄And R ₅In at least one is Cl; And R ₆Be H; Comprise cl anion (that is) reaction that formula 3 compounds and chlorine radical, cl anion or original position are produced such as liquor natrii hypochloritis or hypochlorous acid, from but any R of I or Br ₁, R ₂, R ₃, R ₄Or R ₅Can replace with Cl independently.

Another embodiment relates to formula 3 compounds, wherein is selected from R ₂, R ₃, R ₄Or R ₅At least one but to be no more than two positions are Cl, and the R of each position ₁Be Br, F, Cl or I independently; Condition is not to be any R of Cl or H ₂, R ₃, R ₄Or R ₅Be the R of I and each position ₆Be H or C independently ₁-C ₄Alkyl.

Formula 3 compounds described herein have and are used for characteristic medicine, medical science, the industry of cosmetics and colouring agent.Correspondingly, the compound of the medicine that being used for of the present invention also relates to require to protect uses in part or the body comprises the active substance as the medicine of chemotherapeutic treatment that is used for the mankind or Animal diseases or optical dynamic therapy with it.

The medicine and the purposes of formula 3 compounds, its preparation method, this paper definition should comprise saponification or whole forms of the said compound of reaction in addition, and said reaction is converted into its quinoid (formula 4), wherein R with said compound from its lactone form (formula 3) ₁₁And R ₁₂Be other equilibrium ion that H or Na, K, Li maybe can form salt independently.

In another embodiment, the present invention relates to prepare the method for formula 4 compounds, wherein R ₁Be Cl or Br independently, R ₂, R ₃, R ₄And R ₅Be I, and R ₆Be H.In being substantially free of the solution of cl anion, with formula 3a compound, wherein R ₁Be Cl or Br independently, and R ₂, R ₃, R ₄, R ₅And R ₆Be H, make up to form formula 4 compounds, wherein R with iodide ₁Be Cl or Br independently, R ₂, R ₃, R ₄And R ₅Be I, and R ₆Be H, this provides the essence of the method formerly of relative preparation formula 4 compounds to improve, and said improvement comprises the trans halide derivative that does not have formula 4 compounds basically, wherein R ₁Be Cl or Br independently, R ₂, R ₃, R ₄And R ₅In at least one is Cl and is not any R of Cl ₂, R ₃, R ₄And R ₅Be I, and R wherein ₆Be H.

In another embodiment, the present invention relates to prepare the method for formula 3 compounds,

R wherein ₁Be Cl or Br independently; R ₂, R ₃, R ₄And R ₅Be I; And R ₆Be H, comprise

Step 1: in acid solution, with formula 1 compound,

R wherein ₁Be Cl or Br independently, with the formula 2 compounds combination of about 2 equivalents,

R wherein ₇, R ₈, R ₉And R ₁₀Be H;

Be mixed together in 20 ℃ to 250 ℃ temperature and preferred 85 ℃ of-95 ℃ of temperature; And

The cyclisation intermediate product of separating consequent formula 3a.

R wherein ₁Be Cl or Br and R independently ₂, R ₃, R ₄, R ₅And R ₆Be H; Subsequently

Step 2: with said formula 3a intermediate and not chloride anionic aqueous solution combination;

With the said solution of formula 3a intermediate with iodine (I ₂) handle, mix a period of time 20 ℃ to 100 ℃ temperature, make and accomplish basically to the conversion of formula 3, for example confirm by HPLC or similar means;

The reactant mixture that will contain formula 3 is with the iodine trapping agent quencher that does not conform to cl anion;

Said quencher reactant mixture is acidified to pH less than 5 with not chloride anionic acid solution; And

The said final products of separate type 3.

Also preferred formula 1 and formula 2 compounds do not contain or are substantially free of cl anion and do not contain or be substantially free of the reagent that can in reactant mixture, produce cl anion in the acid solution of the step 1 of combination wherein in the said method of preparation formula 3 compounds.

Also preferred formula 3a intermediate in step 2 does not contain or is substantially free of cl anion and does not contain or be substantially free of reagent or other impurity that can in reactant mixture, produce cl anion in the said method of preparation formula 3 compounds.

Also the solution that makes up therein of preferred formula 3a intermediate and iodine has alkaline pH in the said method of preparation formula 3 compounds.

In the modification of the said method of preparation formula 3 compounds, the step 1 for preparing the method for formula 3 compounds can comprise formula 1 compound and formula 2 compounds combination less than 2 equivalents.This embodiment is not too preferred, and reason is that the yield ratio of formula 3 is lower with the yield that stoichiometry provided of preferred implementation.

R wherein ₁Be I independently, Br, Cl, F, C ₁-C ₄Alkyl or H, and R ₂, R ₃, R ₄And R ₅Be I, F, Cl independently, C ₁-C ₄Alkyl or H, wherein R ₂, R ₃, R ₄Or R ₅In at least one but to be no more than three be I, comprising:

Step 1: not containing or being substantially free of cl anion and not containing or be substantially free of in the acid solution of the reagent that can in reactant mixture, produce cl anion or impurity, with formula 1 compound,

R wherein ₁Be I independently, Br, Cl, F, C ₁-C ₄Alkyl or H and the combination of formula 2 compounds

R wherein ₇, R ₈, and R ₁₀Be F independently, Cl, H, or C ₁-C ₄Alkyl and R ₇, R ₈Or R ₁₀In at least two be H and R ₉Be H;

Temperature at 20 ℃ to 250 ℃ is mixed together;

Separate consequent formula 3a cyclisation intermediate product, for example through filtering or similar means; Subsequently

Step 2: will not contain or be substantially free of cl anion and do not contain or be substantially free of impurity maybe can produce cl anion impurity said formula 3a intermediate with do not contain cl anion or be substantially free of the aqueous solution of cl anion combined;

With the said aqueous solution of formula 3a intermediate with iodine (I ₂) handle;

Mix the sufficient time 20 ℃ to 100 ℃ temperature, make and accomplish basically, for example confirm by HPLC or similar means to the conversion of formula 3;

The reactant mixture that will contain formula 3 is with the quencher of not chloride anionic iodine trapping agent;

Said quencher reactant mixture is acidified to pH＜5 with not chloride anionic acid solution; And

Separate said final products through filtration or similar means.

The said method of preparation formula 3 compounds preferably the aqueous solution of the step 2 that makes up therein of formula 3a intermediate and iodine have alkaline pH.

Further preferably step 2 is mixed the time that continues abundance, makes to accomplish at least 90% and more preferably at least 95% and most preferably at least 98% to the conversion of formula 3, and the said time was generally about 1 to 24 hour and more preferably from about 2 to 18 hours.

In another embodiment, the present invention relates to formula 4 compounds, wherein R ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄, R ₅At least one substituting group be that I and at least one other substituting group are Cl or H; And R ₆Be H or C independently ₁-C ₄Alkyl; And whole (a) tautomeric forms, (b) atropisomer, (c) formula 3 described closed lactone form, (d) enantiomer of formula 3 described lactone form and (e) its pharmaceutically acceptable salt.

Description of drawings

Fig. 1 a illustrates 4,5,6,7-tetrachloro-3 ', and 6 '-dihydroxy-2 ', 4 ', 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,5,6,7-tetrachloro-2 ', 4 ', 5 ', 7 '-tetraiodofluorescein or rose-red lactone;

Fig. 1 b illustrates 4,5,6,7-tetrabromo-3 ', and 6 '-dihydroxy-2 ', 4 ', 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,5,6, the 7-tetrabromobisphenol ', 4 ', 5 ', 7 '-tetraiodofluorescein;

Fig. 1 c illustrates 2 ', 4,5,6,7-pentachloro--3 ', 6 '-dihydroxy-4 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 2 ', 4,5,6,7-pentachloro--4 ', 5 ', 7 '-the triiodo fluorescein;

Fig. 1 d illustrates 4,4 ', 5,6,7-pentachloro--3 ', 6 '-dihydroxy-2 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,4 ', 5,6,7-pentachloro--2 ', 5 ', 7 '-the triiodo fluorescein;

Fig. 1 e illustrates 2 ', 4, and 5,6,7,7 '-chlordene-3 ', 6 '-dihydroxy-4 ', 5 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone are also referred to as 2 ', 4,5,6,7,7 '-chlordene-4 ', 5 '-diiodofluorescein;

Fig. 1 f illustrates 4, and 4 ', 5,5 ', 6,7-chlordene-3 ', 6 '-dihydroxy-2 ', 7 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone are also referred to as 4,4 ', 5,5 ', 6,7-chlordene-2 ', 7 '-diiodofluorescein;

Fig. 1 g illustrates 2 ', 4,5,5 ', 6,7-chlordene-3 ', 6 '-dihydroxy-4 ', 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 2 ', 4,5,5 ', 6,7-chlordene-4 ', 7 '-diiodofluorescein;

Fig. 1 h illustrates 4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,5,6,7-tetrachloro-2 ', 4 ', 5 '-triiodo fluorescein;

Fig. 1 i illustrates 4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,5,6,7-tetrachloro-2 ', 4 ', 7 '-triiodo fluorescein;

Fig. 1 j illustrates 4,5,6, the 7-tetrabromobisphenol '-chloro-3 ', 6 '-dihydroxy-4 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 2 '-chloro-4,5,6,7-tetrabromo-4 ', 5 ', 7 '-triiodo fluorescein;

Fig. 1 k illustrates 4,5,6,7-tetrabromo-4 '-chloro-3 ', 6 '-dihydroxy-2 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4 '-chloro-4,5,6, the 7-tetrabromobisphenol ', 5 ', 7 '-triiodo fluorescein;

Fig. 1 l illustrates 4,5,6, the 7-tetrabromobisphenol ', 7 '-two chloro-3 ', 6 '-dihydroxy-4 ', 5 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 2 ', 7 '-two chloro-4,5,6,7-tetrabromo-4 ', 5 '-diiodofluorescein;

Fig. 1 m illustrates 4,5,6,7-tetrabromo-4 ', 5 '-two chloro-3 ', 6 '-dihydroxy-2 ', 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4 ' 5 '-two chloro-4,5,6, the 7-tetrabromobisphenol ', 7 '-diiodofluorescein;

Fig. 1 n illustrates 4,5,6, the 7-tetrabromobisphenol ', 5 '-two chloro-3 ', 6 '-dihydroxy-4 ', 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 2 ', 5 '-two chloro-4,5,6,7-tetrabromo-4 ', 7 '-diiodofluorescein;

Fig. 1 o illustrates 4,5,6,7-tetrabromo-3 ', 6 '-dihydroxy-2 ', 4 ', 5 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,5,6, the 7-tetrabromobisphenol ', 4 ', 5 '-triiodo fluorescein;

Fig. 1 p illustrates 4,5,6,7-tetrabromo-3 ', 6 '-dihydroxy-2 ', 4 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,5,6, the 7-tetrabromobisphenol ', 4 ', 7 '-triiodo fluorescein;

Fig. 1 q illustrates the atropisomer (aR form) of asymmetric replacement xanthene; With

Fig. 1 r illustrates the atropisomer (aS form) of asymmetric replacement xanthene.

Fig. 1 s illustrates 4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 '-three iodo-7 '-isopropyl-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 7 '-isopropyl-4,5,6,7-tetrachloro-2 ', 4 ', 5 '-triiodo fluorescein.

Fig. 1 t illustrates 2,3,4, and 5-tetrachloro-6-(6-hydroxyl-2,4,5, the 7-tetraiodo-3-oxo-3H-xanthene-9-yl) benzoic acid disodium salt is also referred to as 4,5,6,7-tetrachloro-2 ', 4 ', 5 ', 7 '-tetraiodofluorescein disodium or rose-red.

The detailed description of the preferred embodiment for the present invention

Definition

" C ₁-C ₄Alkyl " is meant the saturated or unsaturated hydrocarbon group of straight chain and branching, generally has the appointment carbon number.C ₁-C ₄The instance of alkyl includes, but not limited to methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, sec-butyl, isobutyl group and the tert-butyl group.

" halo " can use with " halogen " interchangeably, and refers to fluorine, chlorine, bromine and iodine functional group.

" be substantially free of cl anion " and be meant the impurity that reaction condition, reagent or intermediate do not contain cl anion, the impurity that contains cl anion maybe can produce cl anion with similar language; The trans halogenation impurity that its purity level is enough to avoid undesirable formula 3, formula 3a or formula 4 with 0.15% (for example; 1500ppm) or higher level form, perhaps wherein said reaction condition, reagent or intermediate are with 1500ppm or lower level contain cl anion, the impurity that contains cl anion maybe can produce cl anion impurity.

" be substantially free of trans halogenation impurity " and be meant that with similar language formula 3 and formula 4 compounds exist with 0.15% (for example 1500ppm) or lower level, wherein R ₁Be Cl or Br independently, R ₂, R ₃, R ₄And R ₅In at least one is Cl and remaining is I, and R ₆Be H.

General structure is represented

From simplifying reason; The structural table of various intermediates and formula 3, formula 3a, formula 3b and formula 4 final products is shown general lactone or quinoid isomeric form; And depend on that pH or other condition can exist with its tautomeric form (for example, quinoid rather than lactone, or lactone rather than quinoid).Said general expression is not expected disclosure is restricted to illustrational specific general isomeric form.

Reaction scheme

Scheme I shows one embodiment of the present invention, and it involves the method for preparing iodate xanthene (for example formula 3).This method comprises phthalic anhydride (formula 1) and excessive resorcinol (formula 2) reaction, formula 3a is provided compound, wherein R ₂, R ₃, R ₄, R ₅And R ₆Not I and R ₂-R ₅In at least one is H.Then, with the compound iodate, provide formula 3 compounds, wherein R at minimum after separating ₂, R ₃, R ₄Or R ₅In at least one replaces with I.

Scheme I

The cyclization of above-mentioned steps 1 can carry out like this: for example use pure methanesulfonic acid to 10% methanesulfonic acid (MSA) aqueous solution 0 ℃ of temperature to backflow.In preferred embodiment,, carried out 1-16 hour in 85 ℃-95 ℃ pure methanesulfonic acids of use.Another preferred embodiment in, use the MSA of 2-6 volume, more preferably the MSA of 4 to 5 volumes and even the MSA of more preferably about 4.8 volumes.Alternatively, individually or with methanesulfonic acid in combination, can be with p-methyl benzenesulfonic acid; Benzene sulfonic acid, sulfuric acid or TFMS, ethyl sulfonic acid; Acetate, propionic acid, trifluoroacetic acid; Camphorsulfonic acid, or contain fusing point less than about 250 ℃ alkyl sulfonic acid or aryl sulfonic acid, fusing point less than one or more acid solution in the lewis acid of about 250 ℃ alkyl carboxylic acid or aryl carboxylic acid, achloride Bronsted acid, achloride or chloride-immobilization (chloride-immobilized), or their prepared product or salt prefs of polymer-bound separately; Or its aqueous solution, be used for causing cyclisation.

The reactant (for example, 2: 1 formula of ratio, 1 resorcinol and formula 2 phthalic anhydrides) that forms the enough stoichiometric amounts basically of cyclisation ability of formula 3a carries out.In another embodiment, can be preferably with excessive resorcinol (for example, about 2.5 equivalents to about 3 or more equivalents and more preferably from about 3.2 or more equivalents) react so that guarantee the full consumption phthalic anhydride.After separate type 3a, under agitation or not add and can the combination of consequent solid water and acetone and water combination or DMF and water preferably be heated to 50 ℃ to 70 ℃ and more preferably suspend again with acetone and water with stirring, to improve purity at about 60 ℃.Can repeated isolation and suspend again and reach the purity of hope until formula 3a material.

The iodination reaction of preceding text step 2 can be used the excessive iodine (I of molar concentration ₂) preferably under alkali condition, carry out.For example, said reaction can be carried out 0 ℃ of temperature to backflow with 0.1-5M sodium hydroxide, potassium hydroxide, sodium bicarbonate or saleratus.In preferred embodiment, this reaction can be carried out in 70 ℃-95 ℃ with 0.4-1.0M NaOH.Can the mixture of KI, NaI or KI and NaI be used in reactant mixture, for example using the NaI of 1-2.5 equivalent with the iodine solvation.The scope in reaction time be 1-24 hour its depend on the iodine atomicity that is added to the xanthene ring system.

In another embodiment, possibly control the iodate degree through changing reaction time, temperature or alkali concn.The iodine that is used for this reaction can also come original position to produce such as potassium hydrogen peroxymonosulfate or hydrogen peroxide and the iodide salt that is selected from KI, lithium iodide, sodium iodide or its some mixture with oxidant.In this special instance; Preferably avoid to cause the component of undesirable trans halogenation to be incorporated into reactant mixture; Maybe can produce other component of halide ion such as aqueous chloride solution, chlorine, HCl, only if before said component adds, removed or quencher comprises any oxidant of oxygen atmosphere and iodine.For example clorox is undesirable especially component in this reactant mixture; Reason is that it is the source of unstable chlorine anion and effective oxidant, and cl anion, chlorine residue, other hypochlorite derivative, hypochlorous acid and composition thereof also can cause undesirable side reaction in addition.When reaction is accomplished, can mixture be cooled to approximately-20 ℃ to 10 ℃, preferably be lower than 10 ℃, and can add the iodine trapping agent such as sodium thiosulfate, potassium thiosulfate, ATS (Ammonium thiosulphate), potassium sulfite, sodium sulphite or its mixture quencher iodine.In preferred embodiment, carry out quencher with sodium sulphite.

It is possible that the formula of being easy to 3 is separated: preferably keep temperature to be lower than 10 ℃ also with pure aqueous sulfuric acid to 1% aqueous sulfuric acid, and preferred 5% aqueous sulfuric acid conditioned reaction pH, to 5, this causes formula 3 self-dissolving liquid precipitates to come out until pH regulator to 1.5.In preferred embodiment, with pH regulator to pH 1.5 to pH 3.After separate type 3, can consequent solid be suspended again, and in for example water and the combination of acetone and water or the combination of DMF and water of environmental temperature, the combination of preferred acetone and water further separates to remove impurity.Extraly, can pH regulator to 5 or higher so that formula 3 solvents are turned to formula 4 be transformed back formula 3 subsequently so that under acid condition, separate.

Scheme II

Scheme II shows another embodiment of the invention, and its Chinese style 3 compounds can also exist with quinoid form (formula 4) in basic neutrality or alkaline pH, and these quinoids can be used as salt existence, wherein R ₁₁And/or R ₁₂One or two hydroxyl with comprising Na, the replacement of the alkaline equilibrium ion of K or Li ion.

Scheme III has shown the formula of starting from 3 compounds, wherein is selected from R ₂, R ₃, R ₄And R ₅At least one position be I, with the method for Cl substitute I, wherein be selected from R with production 3b compound ₂, R ₃, R ₄And R ₅At least one position be Cl.

Scheme III

Scheme IV shows another embodiment of the invention, and its Chinese style 4 compounds can also separate as the product of this process, and these quinoids can be used as salt existence, wherein R ₁₁And/or R ₁₂One or two hydroxyl with the equilibrium ion replacement that can form pharmaceutically acceptable salt that comprises H, Na, K or Li ion.

Scheme IV

Preferred implementation of the present invention relates to formula 3 and formula 4 compounds, wherein R ₁Be Cl or Br independently, and R ₂-R ₅Be to be selected from Cl, I or H independently, wherein at least 3 hydrogen atoms are present in R ₂-R ₆Or at least one chlorine atom is present in R ₂-R ₅, and comprise atropisomer when feasible.

One group of embodiment of the present invention comprises following compound and pharmaceutically acceptable salt thereof, and called after lactone form (formula 3) and quinoid form (formula 4), like the explanation of Fig. 1:

4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 '; 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,5,6,7-tetrachloro-2 ', 4 '; 5 ', 7 '-tetraiodofluorescein, referring to Fig. 1 a, and isomery quinoid form 2,3,4,5-tetrachloro-6-(6-hydroxyl-2; 4,5, the 7-tetraiodo-3-oxo-3H-xanthene-9-yl) benzoic acid, referring to Fig. 1 t, be commonly referred to as rose-red, the perhaps Sodium tetraiodotetrachlorofluorescein of its disodium salt form;

4,5,6,7-tetrabromo-3 ', 6 '-dihydroxy-2 ', 4 '; 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,5,6; The 7-tetrabromobisphenol ', 4 ', 5 ', 7 '-tetraiodofluorescein, referring to Fig. 1 b, and isomery quinoid form 2; 3,4,5-tetrabromo-6-(6-hydroxyl-2,4,5, the 7-tetraiodo-3-oxo-3H-xanthene-9-yl) benzoic acid;

2 ', 4,5,6,7-pentachloro--3 ', 6 '-dihydroxy-4 ', 5 '; 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 2 ', 4,5,6,7-pentachloro--4 '; 5 ', 7 '-the triiodo fluorescein, referring to Fig. 1 c, and isomery quinoid form 2,3,4,5-tetrachloro-6-(2-chloro-6-hydroxyl-4; 5,7-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid and 2,3,4,5-tetrachloro-6-(7-chloro-6-hydroxyl-2,4,5-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid;

4,4 ', 5,6,7-pentachloro--3 ', 6 '-dihydroxy-2 ', 5 '; 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,4 ', 5,6,7-pentachloro--2 '; 5 ', 7 '-the triiodo fluorescein, referring to Fig. 1 d, and isomery quinoid form 2,3,4,5-tetrachloro-6-(4-chloro-6-hydroxyl-2; 5,7-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid and 2,3,4,5-tetrachloro-6-(5-chloro-6-hydroxyl-2,4,7-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid;

2 ', 4,5,6,7,7 '-chlordene-3 '; 6 '-dihydroxy-4 ', 5 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone are also referred to as 2 ', 4,5; 6,7,7 '-chlordene-4 ', 5 '-diiodofluorescein, referring to Fig. 1 e, and quinoid form 2; 3,4,5-tetrachloro-6-(2,7-two chloro-6-hydroxyls-4,5-two iodo-3-oxos-3H-xanthene-9-yl) benzoic acid;

4,4 ', 5,5 ', 6,7-chlordene-3 '; 6 '-dihydroxy-2 ', 7 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone are also referred to as 4,4 ', 5; 5 ', 6,7-chlordene-2 ', 7 '-diiodofluorescein, referring to Fig. 1 f, and quinoid form 2; 3,4,5-tetrachloro-6-(4,5-two chloro-6-hydroxyls-2,7-two iodo-3-oxos-3H-xanthene-9-yl) benzoic acid;

2 ', 4,5,5 ', 6,7-chlordene-3 ', 6 '-dihydroxy-4 '; 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 2 ', 4,5,5 ', 6; 7-chlordene-4 ', 7 '-diiodofluorescein, referring to Fig. 1 g, and isomery quinoid form 2,3,4,5-tetrachloro-6-(2; 5-two chloro-6-hydroxyls-4,7-two iodo-3-oxos-3H-xanthene-9-yl) benzoic acid and 2,3,4,5-tetrachloro-6-(4,7-two chloro-6-hydroxyls-2,5-two iodo-3-oxos-3H-xanthene-9-yl) benzoic acid;

4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 '-three iodo-3H-spiral shell [isobenzofurans-1; 9 '-xanthene]-3-ketone, be also referred to as 4,5,6,7-tetrachloro-2 ', 4 ', 5 '-triiodo fluorescein; Referring to Fig. 1 h, and isomery quinoid form 2,3,4,5-tetrachloro-6-(6-hydroxyl-2,4; 5-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid and 2,3,4,5-tetrachloro-6-(6-hydroxyl-4,5,7-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid;

4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 7 '-three iodo-3H-spiral shell [isobenzofurans-1; 9 '-xanthene]-3-ketone, be also referred to as 4,5,6,7-tetrachloro-2 ', 4 ', 7 '-triiodo fluorescein; Referring to Fig. 1 i, and isomery quinoid form 2,3,4,5-tetrachloro-6-(6-hydroxyl-2,4; 7-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid and 2,3,4,5-tetrachloro-6-(6-hydroxyl-2,5,7-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid;

4,5,6, the 7-tetrabromobisphenol '-chloro-3 ', 6 '-dihydroxy-4 ', 5 '; 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 2 '-chloro-4,5,6,7-tetrabromo-4 '; 5 ', 7 '-triiodo fluorescein, referring to Fig. 1 j, and isomery quinoid form 2,3,4; 5-tetrabromo-6-(2-chloro-6-hydroxyl-4,5,7-three iodo-3-oxos-9,9a-dihydro-3H-xanthene-9-yl) benzoic acid and 2,3; 4,5-tetrabromo-6-(7-chloro-6-hydroxyl-2,4,5-three iodo-3-oxos-9,9a-dihydro-3H-xanthene-9-yl) benzoic acid;

4,5,6,7-tetrabromo-4 '-chloro-3 ', 6 '-dihydroxy-2 ', 5 '; 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4 '-chloro-4,5,6, the 7-tetrabromobisphenol '; 5 ', 7 '-triiodo fluorescein, referring to Fig. 1 k, and isomery quinoid form 2,3,4; 5-tetrabromo-6-(4-chloro-6-hydroxyl-2,5,7-three iodo-3-oxos-9,9a-dihydro-3H-xanthene-9-yl) benzoic acid and 2,3; 4,5-tetrabromo-6-(5-chloro-6-hydroxyl-2,4,7-three iodo-3-oxos-9,9a-dihydro-3H-xanthene-9-yl) benzoic acid;

4,5,6, the 7-tetrabromobisphenol ', 7 '-two chloro-3 ', 6 '-dihydroxy-4 '; 5 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 2 ', 7 '-two chloro-4,5,6; 7-tetrabromo-4 ', 5 '-diiodofluorescein, referring to Fig. 1 l, and quinoid form 2,3; 4,5-tetrabromo-6-(2,7-two chloro-6-hydroxyls-4,5-two iodo-3-oxos-9,9a-dihydro-3H-xanthene-9-yl) benzoic acid;

4,5,6,7-tetrabromo-4 ', 5 '-two chloro-3 ', 6 '-dihydroxy-2 '; 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4 ' 5 '-two chloro-4,5,6; The 7-tetrabromobisphenol ', 7 '-diiodofluorescein, referring to Fig. 1 m, and quinoid form 2,3; 4,5-tetrabromo-6-(4,5-two chloro-6-hydroxyls-2,7-two iodo-3-oxos-9,9a-dihydro-3H-xanthene-9-yl) benzoic acid;

4,5,6, the 7-tetrabromobisphenol ', 5 '-two chloro-3 ', 6 '-dihydroxy-4 '; 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 2 ', 5 '-two chloro-4,5,6; 7-tetrabromo-4 ', 7 '-diiodofluorescein, referring to Fig. 1 n, and isomery quinoid form 2,3,4; 5-tetrabromo-6-(2,5-two chloro-6-hydroxyls-4,7-two iodo-3-oxos-9,9a-dihydro-3H-xanthene-9-yl) benzoic acid and 2,3; 4,5-tetrabromo-6-(4,7-two chloro-6-hydroxyls-2,5-two iodo-3-oxos-9,9a-dihydro-3H-xanthene-9-yl) benzoic acid;

4,5,6,7-tetrabromo-3 ', 6 '-dihydroxy-2 ', 4 ', 5 '-three iodo-3H-spiral shell [isobenzofurans-1; 9 '-xanthene]-3-ketone, be also referred to as 4,5,6, the 7-tetrabromobisphenol ', 4 ', 5 '-triiodo fluorescein; Referring to Fig. 1 o, and isomery quinoid form 2,3,4,5-tetrabromo-6-(6-hydroxyl-2,4; 5-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid and 2,3,4,5-tetrabromo-6-(6-hydroxyl-4,5,7-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid;

4,5,6,7-tetrabromo-3 ', 6 '-dihydroxy-2 ', 4 ', 7 '-three iodo-3H-spiral shell [isobenzofurans-1; 9 '-xanthene]-3-ketone, be also referred to as 4,5,6, the 7-tetrabromobisphenol ', 4 ', 7 '-triiodo fluorescein; Referring to Fig. 1 p, and isomery quinoid form 2,3,4,5-tetrabromo-6-(6-hydroxyl-2,4; 7-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid or isomer 2,3,4,5-tetrabromo-6-(6-hydroxyl-2,5,7-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid;

Under the asymmetric substituted situation of xanthene core, the aR form of atropisomer referring to Fig. 1 q and aS form, referring to Fig. 1 r, is possible at one of these compounds; And

4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 '; 5 '-three iodo-7 '-isopropyl-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone, be also referred to as 4,5,6; 7-tetrachloro-7 '-isopropyl-2 ', 4 ', 5 '-triiodo fluorescein, referring to Fig. 1 s, and isomery quinoid form 2; 3,4,5-tetrachloro-6-(7-isopropyl-6-hydroxyl-2,4,5-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid and 2; 3,4,5-tetrachloro-6-(2-isopropyl-6-hydroxyl-4,5,7-three iodo-3-oxos-3H-xanthene-9-yl) benzoic acid.

Formula 3 of the present invention or formula 4 compounds can contain chiral centre, and therefore can exist with different enantiomeric forms and diastereomer form.The present invention relates to whole optical isomers and whole stereoisomers as racemic mixture and the independent enantiomer of conduct of formula 3 or formula 4 compounds; Diastereoisomer with said compound; And composition thereof, and relate to the whole pharmaceutical compositions that contain them of this paper definition or their methods of treatment of application of this paper definition.In preparation final products or its intermediate, individual isomers can obtain such as optical resolution, fractional crystallization, optical selective reaction condition or chromatographic isolation through known method.

Similarly, the quinoid form of formula 3 or formula 4 compounds, wherein said xanthene ring since corresponding xanthene aryl rings on different substituents (also promptly for example, at R ₃=R ₄Situation under R ₂≠ R ₅Or at R ₂=R ₅Situation under R ₃≠ R ₄, such as the quinoid form of compound described in Fig. 1 c, 1d, 1g, 1h, 1i, 1j, 1k, 1o, the 1p) and it is asymmetric, can be used as and stablize atropisomer and have () of Fig. 1 q and 1r.The racemic atropisomer can be used as that reaction impurities exists and situation as these atropisomers of racemic mixture preparation under, process described herein contains and minimizes the right relative quantity of racemic atropisomer.

At formula 3 compounds is under the situation of acid compound, and they can form different salt miscellaneous with various inorganic and organic bases.The base addition salts of acid compound of the present invention prepares through formula 3 lactones being handled easily with the selected inorganic base of 1 or 2 equivalents or organic base in such as ethanol or methyl alcohol at aqueous solvent or suitable organic solvent at least.After evaporating solvent, filtration or directly using the aqueous solution of gained salt, desirable salt is described suc as formula 4 and is easily obtained with the quinoid form.Pharmaceutically acceptable salt for example comprises those that form with sodium, calcium, potassium, magnesium, meglumine, ammonium, aluminium, zinc, piperazine, tromethamine, lithium, choline, diethylamine, 4-benzyl ring hexylamine and N, N '-dibenzyl-ethylenediamin.

The present invention such as also comprises at those compound isotopically labelled of cotype 3 and formula 4, except one or more atoms are different from atomic mass or the atomic mass of mass number or the atom replacement of mass number that natural world exists usually with having.Can mix isotope that isotopic instance in the The compounds of this invention comprises hydrogen, carbon, oxygen, fluorine, chlorine and iodine respectively such as ²H, ³H, ¹⁴C, ¹³C, ¹⁰C, ¹¹C, ¹³O, ¹⁴O, ¹⁵O, ¹⁸O, ¹⁷O, ¹⁷F, ¹⁸F, ³²Cl, ³³Cl, ³⁴Cl, ³⁶Cl, ⁷⁴Br, ⁷⁵Br, ⁷⁶Br, ⁷⁷Br, ¹¹⁷I, ¹¹⁸I, ¹²⁰I, ¹²¹I, ¹²²I, ¹²⁴I, ¹²⁶I, ¹²⁸I with ¹³¹I.Contain other its prodrug of isotopic The compounds of this invention of aforementioned isotope and/or other atom and the pharmaceutically acceptable salt of said compound or the pharmaceutically acceptable salt of said prodrug and belong to scope of the present invention.Some compound isotopically labelled useful as drug distribution tests of the present invention/or substrate organize the diagnosticum of distribution tests.Isotope-labeled formula 3 generally can prepare through the program of implementing to be disclosed among hereinafter scheme and/or the embodiment with formula 4 compounds and prodrug thereof, but replaces nonisotopically labelled reagent with the isotope labeling reagent that obtains easily, also promptly uses ¹³¹I replaces nonisotopically labelled iodine.

The situation of one embodiment of the present invention be formula 4 compounds account for medicine about 0.001% to less than about 20% weight.

One embodiment of the present invention relate to some medical usage of medicine and said medicine and use said medicine to treat the treatment of diseases method of the mankind or animal tissue, and the main active component of wherein said medicine is the halogenation xanthene of formula 3 or formula 4.Said medicine can for example serve as chemical ablation agent or pdt agent on chemotherapy; And can be used for treating various illnesss; Said illness influences skin and relevant organ, oral cavity and digestive tract and relevant organ, urethra and genital tract and relevant organ, respiratory tract and relevant organ, the circulatory system and relevant organ, head and neck, endocrine system and lymphoreticular system and relevant organ, various other tissue; Such as the connective tissue and the various tissue surface that expose at intra-operative, and the various tissues of showing microorganism, virus, fungi or parsitism.

These medicines can obtain with various preparatons, and it can comprise liquid, semisolid, solid or aerosol delivery vector, and is suitable for carrying out vivo medicine-feeding via various normal modes and approach; Said pattern and approach comprise intravenous injection (i.v.), intraperitoneal injection (in the peritonaeum), intramuscular injection (i.m.); Intracranial injection (i.c.), intratumor injection (i.t.), intralesional injection (i.l.); Last intracutaneous injection (also being), dermal delivery (t.c.) and through oesophagus (p.o.) administration.Extraly, said medicine is suitable for via various normal modes and approach topical, and said pattern and approach comprise near direct local application to some tissue or its.Said active ingredient of drugs produces the therapeutic response of hoping; Infect such as destroy microorganisms; Reduce or eliminate tissue stimulation or inflammation, reduce or eliminate the hyper-proliferative tissue, reduce or eliminate cancerous tissue or pre-cancer tissue; Reduce or eliminate surface or surface lipocyte or lipidosis down, with many other similar indications.

In preferred embodiment, said medicine prepares with various preparatons, comprises liquid, semisolid, and solid or aerosol delivery vector, and with tablet, capsule, suppository and the preparation of other similar form.

Another preferred embodiment in, with at least one targeting moiety at R ₁To R ₆The optional position or be coupled to the halogenation xanthene of formula 3 or formula 4 via the connection on hydroxyl or carbonyl.Said targeting moiety can be selected from and include but not limited to following group: DNA (DNA), ribonucleic acid (RNA), amino acid, albumen, antibody, part, hapten; Carbohydrate receptor, carbohydrate compounding ingredient, lipid acceptor, lipid compounding ingredient, protein acceptor, protein compounding ingredient, chelating agent; Encapsulation vehicle, short chain aliphatic hydrocarbon, long chain aliphatic hydrocarbon, aromatic hydrocarbons, aldehyde, ketone, alcohol; Ester, acid amides, amine, nitrile, azide, hydrophilic segment and hydrophobic part.

Another preferred embodiment in, formula 4 compounds can be used in the preparation medicine.

Embodiment

Following embodiment is contemplated to be illustrative rather than restrictive, and represents embodiment of the present invention.

Conventional method

Total overall reaction is carried out in open-top receptacle.Sometimes covering container is to avoid ambient lighting.Total overall reaction is carried out under nitrogen, argon or other inert atmosphere, unless otherwise.Used whole solvents and reagent is from commercial source, and is not further purified.With high pressure liquid chromatography (HPLC) monitoring reaction, it uses Agilent 1100 serial quaternary pump/variable-wavelenght detectors (225nm detects wavelength), and (4.6x150mm, 2.7 μ m be in 40 ℃, MeCN/0.5%H to be furnished with Supelco Ascentis Express C18 post ₃PO ₄H ₂O solution, move 30 minutes in 65/35 to 90/10 gradient), perhaps identical device is furnished with Waters Symmetry Shield RP-18 post (4.6x150mm, 5 μ m is in 40 ℃, MeCN/10mM K ₃PO ₄H ₂O solution (pH 3)+5%MeCN, 10/90 to 80/20 gradient or degree such as 65/35 in 25 minutes); Mass spectrum (Agilent LC/MSD trap or Waters LCMS have the similar chromatography condition of HPLC, modify with formic acid); And/or thin-layer chromatography (TLC), it carries out visual with ultraviolet light and iodine staining.Carry out supercritical fluid chromatography (SFC) with Thar SFC 80, be furnished with RegisCell post (3x 25cm), ethanol carrying capacity 20-22mg/ injection, concentration 4.5mg/mL.In 300MHz VarianINOVA 300 or Varian Gemini 2000 or in 400MHz Varian Oxford 400 record proton magnetic resonance (PMR)s ( ¹H NMR) spectrogram, mark in each personal TMS does.Relative DMSO-d ₆2.50ppm unimodal, with reference to the tetramethylsilane (TMS) of 0ppm, the report chemical shift (δ, unit 1,000,000/, ppm).Coupling constant (J) is pressed Hertz (Hz) report.In 75MHz Varian INOVA 300 or in 100MHz at Varian Oxford 400, or in 75MHz on Varian Gemini 2000 spectrometers, write down noise-decouple 13C-NMR ( ¹³C NMR) spectrogram.Relative DMSO-d ₆Heptet center line in 39.5ppm is reported chemical shift δ with ppm.The UV-VIS spectroscopic data obtains at Hitachi U-2810 Double Beam spectrophotometer or on Spectronic Genesys 2 spectrophotometers, from 200-600nm scanning, slit width 1.5nm, light path 10.0mm.

Embodiment 1-preparation 4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone

(rose-red lactone, Fig. 1 a)

Step 1: cyclisation.The 500mL round-bottomed flask is equipped with heating jacket, J-Kem thermocouple, big magnetic agitation, nitrogen arrival line, and magnetic stirring apparatus.To this equipment add tetrachlorophthalic tetrachlorophthalic anhydrid (1.00 equivalents, 50.00g, 174.9mmol), resorcinol (2.10 equivalents, 40.44g, 367.3mmol) and pure methanesulfonic acid (250mL).The gained reactant mixture at room temperature is a suspension.Reactant mixture purges with nitrogen and is heated to 90 ℃, and dark reddish orange solution is provided.To be reflected at 90 ℃ and keep 5 hours, add then extra 5.78g resorcinol (0.3 equivalent, 52.47mmol).Continued stirring reaction 2.5 hours in 90 ℃.Reaction is regarded as accomplishing under the situation of measuring remaining tetrachlorophthalic tetrachlorophthalic anhydrid amount＜1.0% through HPLC.1L round-bottomed flask (3 neck) is equipped with, mechanical agitator, ice bath and J-Kem thermocouple add environment USP water (500mL).With the nitrogen of normal pressure, 90 ℃ of reactant mixtures are slowly added＜10 ℃ USP water via line of transference.During reactant mixture was transferred to water, the control transfer rate made the temperature of water quencher mixture be no more than 60 ℃.100mL USP water with extra washes reactor, is transferred to the reactor that contains this water then.Let consequent green and brown look suspension be cooled to room temperature (RT) gradually, stirred extra 30 minutes at RT.Via the isolated by vacuum filtration solid.Reactor is washed with USP water (2x 250mL aliquot), and wash wet cake with these flushing liquors.Wet cake was drained 60 minutes,, the green and brown look solid of 88.60g is provided then in a vacuum in 70 ℃ of dried overnight.This material is added the 2L round-bottomed flask with 550mL acetone, the Y adapter that it is furnished with mechanical agitator, J-Kem thermocouple, heating jacket, nitrogen arrival line and is equipped with reflux condenser.Gained suspension be heated to reflux continue 1.5 hours, then with it with 530mL USP water treatment (in 60 minutes, slowly adding) via dropping funel, make＞56 ℃ of temperature maintenances.During adding water, observe temperature rising (maximum temperature that is realized is 62 ℃).After adding the water completion, faint yellow-brown suspension is remained on following 3 hours of backflow, in about 30-40 minute, be cooled to RT gradually then.At room temperature mixture was stirred extra 30 minutes, collect faint yellow-brown suspension via vacuum filtration then.Reactor with 50% aqueous acetone (v/v, 4x 100mL) flushing, is washed wet cake with these flushing liquors.Wet cake being pumped to dried in 48 hours, then further in a vacuum in 70 ℃ of dried overnight, 74.43g being provided the tetrachlorofluorescein of (90.5% yield, 96.8%AUC purity), is the yellowish-brown solid. ¹H NMR (300MHz; DMSO d ₆) δ 10.23 (S, 2H), 6.95 (d, J=8.7Hz, 2H), 6.69 (d, J=2.1Hz, 2H), 6.57 (double doublet, J=8.7Hz, 2H).

Step 2: iodate.500mL round-bottomed flask (3-neck) is equipped with reflux condenser, Y adapter, J-Kem thermocouple, mechanical agitator, heating jacket and nitrogen arrival line.Reactor is purged with nitrogen, cover with aluminium foil.This adds 10.00 gram tetrachlorofluoresceins (21.3mmol), and 30mL 5M NaOH solution and 300mL USP water provide dark red solution.Then, the iodine (127.7mmol) with 7.03 gram sodium iodides (46.9mmol) and 32.4g adds to reactor.Let reactant mixture stir 30 minutes in environmental temperature, be heated to 90 ℃ then.The HPLC of reactant mixture aliquot analyzes and points out the raw material full consumption, does not have the intermediate of part iodate, and is converted into desirable product fully.React on 90 ℃ the heating 1.5 hours after, close thermal source, let reactant mixture in 1.5 hours, be cooled to room temperature gradually.With ice bath darkviolet-pink colour reactant mixture is cooled to＜10 ℃.Reactant mixture pH is 7.13.Divide in small batches sodium sulphite (6.70g) is added reactant mixture.At＜10 ℃ 75mL acetone is added to reactor, let mixture stir 10 minutes at＜10 ℃.During＜10 ℃, Dropwise 5 %H ₂SO ₄The aqueous solution (24mL) is realized pH2.03, produces pink colour suspension.Collect this reaction suspension via vacuum filtration.Reactor with 25% aqueous acetone (v/v, 4x 100mL) flushing, is washed wet cake with flushing liquor.Wet cake was drained 3 hours,, the pink solid of 25.81g is provided in a vacuum in 60 ℃ of dried overnight.These solids and 225mL acetone are added 1L round-bottomed flask (3 neck), and it is furnished with mechanical agitator, the Y adapter of J-Kem thermocouple and tool nitrogen inlet.At room temperature stirred this mixture 10 minutes, and in 10 minutes, added 255mL USP water then, suspension is provided.At room temperature stirred suspension is 2.25 hours, comes the isolated by filtration solid via vacuum filtration then.Reactor is washed with 50% aqueous acetone (1x 75mL), and flushing liquor is used for washing wet cake.Then, wet cake with 50% aqueous acetone (2x 75mL) and USP water (1x 75mL) flushing, was drained 1 hour, further dry in 80 ℃; Provide 18.68g product (90.2% yield, coral pink solid, HPLC AUC purity 99.5%) 4,5; 6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 '; 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone.

¹H?NMR(300MHz；DMSO?d ₆)δ10.27(s，2H)，7.59(s，2H)。 ¹³C?NMR(300MHz；DMSO?d ₆)δ163.2，158.6，151.9，146.7，138.8，136.5，135.3，131.2，126.8，124.6，110.8，81.8，79.3，77.0。MS MSD trap: m/z 974.8 (M+1) ⁺(accurate mass 973.67).UV-VIS λ max=557nm is in the methyl alcohol; The definite fusing point (mp)=215 of differential scanning calorimetry (DSC) ℃.

Embodiment 2-preparation 4,5,6,7-tetrabromo-3 ', 6 '-dihydroxy-2 ', 4 ', 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone

(Fig. 1 b)

Step 1: according to the program of embodiment 1 step 1 description; In the 143mL methanesulfonic acid, handle tetrabromophthalic anhydride (28.63g, 61.74mmol) and resorcinol (17g; 154.4mmol); The eosin of 33.64g is provided, leaves (84.1% yield, AUC purity 97.4%) as shallow rice white solids. ¹H NMR (300MHz; DMSO d ₆) δ 10.17 (s, 2H), 6.86 (d, J=8.64Hz, 2H), 6.65 (d, J=2.34Hz, 2H), 6.54 (double doublet, J=8.7Hz, 2H).

Step 2: according to the program that embodiment 1 step 2 is described, handle eosin (22g, 33.96mmol), iodine (51.71g, 203.7mmol) and sodium iodide (11.22g; 74.85mmol), 4,5 of 36.02g (31.2mmol, 92.1% yield, AUC purity 96.3%) is provided; 6,7-tetrabromo-3 ', 6 '-dihydroxy-2 ', 4 '; 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone is as being that the lightpink solids leaves. ¹H?NMR(300MHz；DMSO?d ₆)δ10.22(s，2H)，7.49(s，2H)。 ¹³C?NMR(300MHz；DMSO?d ₆)δ163.9，158.8，152.5，149.9，137.3，136.7，133.5，127.5，124.6，121.1，111.5，82.0，80.3，77.4。MS MSD trap: m/z 1152.6 (M+1) ⁺(accurate mass 1151.48).UV-VIS λ max=558nm, in the methyl alcohol, mp (DSC mensuration)=227 ℃.

Embodiment 3-preparation 2 ', 4,5,6,7-pentachloro--3 ', 6 '-dihydroxy-4 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone

(Fig. 1 c)

Step 1: according to the program that embodiment 1 step 1 is described, the processing tetrachlorophthalic tetrachlorophthalic anhydrid (5g, 17.48mmol), 4-chloro resorcinol (2.78g; 19.23mmol) and resorcinol (2.12g 19.23mmol), provides 18.6% pure 2 ', 4 of 7.24g; 5,6,7-pentachloro--3 '; 6 '-dihydroxy-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone, be used for subsequent step with its collection and as mixture.

Step 2: according to the program that embodiment 1 step 2 is described, iodate 2 ', 4,5,6,7-pentachloro--3 ', 6 '-dihydroxy-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone (7.24g, 15.4mmol).Then, via SFC purifying (80g/min total flow, 40% cosolvent (EtOH/0.5%TFA)/CO ₂In, 140 crust, 254nm is on 3x25cm 5m RegisPack post) separation coarse products (5.47g); 2 of 427mg (0.54mmol, 34.7% yield) ' is provided, 4,5; 6,7-pentachloro--3 ', 6 '-dihydroxy-4 ', 5 '; 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be the lightpink solid, 95.8%AUC HPLC purity. ¹H?NMR(300MHz；DMSO?d ₆)δ10.85(s，1H)，10.2(s，1H)，7.60(s，1H)，7.38(s，1H)。 ¹³C?NMR(300MHz；DMSO?d ₆)δ163.4，159.0，155.8，152.3，151.2，146.9，139.1，136.9，135.7，131.6，128.2，127.1，125.1，116.9，111.0，109.7，82.3，80.0，78.4，78.4，77.5。MS MSD trap: m/z 882.7 (M+1) ⁺(accurate mass 882.2).UV-VIS λ max=554nm is in the methyl alcohol.

Embodiment 4-is 4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 '; 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone is degraded to 4,4 ', 5,6,7-pentachloro--3 '; 6 '-dihydroxy-2 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone and 4,4 ', 5; 5 ', 6,7-chlordene-3 ', 6 '-dihydroxy-2 ', 7 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone

(Fig. 1 a, 1d and 1f)

At room temperature, to 4,5,6 of 100mg (0.10mmol), 7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone adds 1.0mL acetonitrile and 2.0mL 12.5% moisture clorox.At RT reactant mixture was stirred 1 hour.HPLC analyze point out in 8.38 minutes with two kinds of new impurity of 9.93 minutes retention times (respectively 27.8% and 45.6%), remain 26.6% 4,5,6; 7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 '; 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone unreacted.Mass spectral analysis shows M-1 ion: M-1=791.7 corresponding to chlordene impurity, M-1=881.4 corresponding to pentachloro-impurity and M-1=973.2 corresponding to rose-red lactone.Hereinafter prepares the compound (referring to embodiment 5 and 7) of trans halogenation individually, to confirm architectural feature and to compare with HPLC and this embodiment product.

Embodiment 5-preparation 4,4 ', 5,6,7-pentachloro--3 ', 6 '-dihydroxy-2 ', 5 ', 7 '-three iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone

(Fig. 1 d)

Step 1: with tetrachlorophthalic tetrachlorophthalic anhydrid (2.5g; 8.74mmol), the 2-chloro resorcinol (1.33g, 9.18mmol) and resorcinol (1.01g; 9.18mmol) combined with the pure methanesulfonic acid of 12.5mL; Be heated to 90 ℃ and continue 19 hours, continue 10 hours to 97 ℃ then, then hot mixt is carefully added the 25mL frozen water.This suspension is extracted into ethyl acetate, water and brine wash, dry on sodium sulphate then.Product via silicagel pad with 66%: 14%: 18%: 4% toluene ∶ diox: hexane: acetate separates as eluent.Collect 1.95g 46.4% pure 4,4 ', 5,6,7-pentachloro--3 ', 6 '-dihydroxy-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone, do not add and be used for subsequent step with being further purified.

Step 2: according to the program that embodiment 1 step 2 is described, iodate 4,4 ', 5,6,7-pentachloro--3 ', 6 '-dihydroxy-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone (1.95g, 4.15mmol).The mixture of rough acquisition 2.70g.Via SFC purifying (80g/min total flow, 40% cosolvent IPA/CO ₂, 140 crust, 254nm is on 3x25cm 5m RegisPack post) products of separated; 4,4 ' of 630mg (0.7mmol, 38.5% yield, 98.7%AUC purity) is provided; 5,6,7-pentachloro--3 ', 6 '-dihydroxy-2 '; 5 ', 7 '-three iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone are pink solid. ¹H?NMR(300MHz；DMSO?d ₆)δ11.05(s，1H)，10.3(s，1H)，8.3(s，1H)，7.55(d，2H)。 ¹³C?NMR(300MHz；DMSO?d ₆)δ163.2，158.6，154.7，150.7，148.0，146.7，138.7，136.5，135.3，134.5，131.3，126.8，124.6，110.5，110.4，82.4，82.1，79.3，77.6。MS MSD trap: m/z 882.7 (M+1) ⁺(accurate mass 882.2).UV-VIS λ max=555nm is in the methyl alcohol.

Embodiment 6-preparation 2 ', 4,5,6,7,7 '-chlordene-3 ', 6 '-dihydroxy-4 ', 5 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone

(Fig. 1 e)

Step 1: according to the program that embodiment 1 step 1 is described, (5g, 17.49mmol) (6.32g 43.73mmol), provides 10.31g coarse products with the 4-chloro resorcinol to handle tetrachlorophthalic tetrachlorophthalic anhydrid.9.25g part the said goods is suspended in DMF: in the water (1: 1), subsequent filtration provides 9.70g (2: 1DMF compound 17.16mmol; 92.3% yield) 2 ', 4,5; 6,7,7 '-chlordene-3 '; 6 '-dihydroxy-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone, separate as orange solids. ¹HNMR(300MHz；DMSO?d ₆)δ11.13(S，2H)，7.31(s，2H)，6.89(s，2H)。

Step 2:, handle 2 ', 4,5,6,7 according to the program that embodiment 1 step 2 is described; 7 '-chlordene-3 ', 6 '-dihydroxy-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-(2g 3.71mmol), provides 2 ' of 2.56g (3.22mmol, 87.3% yield) to 3-ketone; 4,5,6,7,7 '-chlordene-3 '; 6 '-dihydroxy-4 ', 5 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone separate 97.9%AUC HPLC purity as the powder orange solids. ¹H?NMR(300MHz；DMSO?d ₆)δ10.90(s，2H)，7.48(s，2H)。 ¹³C?NMR(300MHz；DMSO?d ₆)δ163.2，154.7，147.1，146.8，138.7，135.3，134.5，131.3，126.7，124.5，110.3，107.7，82.6，79.0。MS MSD trap: m/z 791.0 (M+1) ⁺(accurate mass 790.77).UV-VIS λ max=551nm is in the methyl alcohol.

Embodiment 7-preparation 4,4 ', 5,5 ', 6,7-chlordene-3 ', 6 '-dihydroxy-2 ', 7 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone

(Fig. 1 f)

Step 1: according to the program that embodiment 1 step 1 is described, (5g, 17.49mmol) (7.58g 41.98mmol), provides the coarse products of 9.3g with the 2-chloro resorcinol to handle tetrachlorophthalic tetrachlorophthalic anhydrid.8g in the 9.3g coarse products is suspended in DMF: in the water (1: 1), subsequent filtration provides 4 of 8.65g (16.05mmol, 91.7%AUC yield); 4 ', 5,5 ', 6; 7-chlordene-3 ', 6 '-dihydroxy-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone, separate as orange solids. ¹H?NMR(300MHz；DMSO?d ₆)δ11.09(S，2H)，7.03(d，J＝9Hz，2H)，6.81(d，J＝8.7Hz，2H)。

Step 2: according to the program that embodiment 1 step 2 is described, processing 4,4 ', 5,5 ', 6; 7-chlordene-3 ', 6 '-dihydroxy-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-(2g 3.71mmol), provides 4 of 2.20g (2.78mmol, 75.1% yield) to 3-ketone; 4 ', 5,5 ', 6,7-chlordene-3 '; 6 '-dihydroxy-2 ', 7 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone separate 97.7%AUC HPLC purity as pink colour-orange solids. ¹H?NMR(300MHz；DMSO?d ₆)δ11.05(s，2H)，7.58(s，2H)。 ¹³C?NMR(300MHz；DMSO?d ₆)δ163.2，154.7，147.1，146.8，138.7，135.3，134.5，131.3，126.7，124.5，110.3，107.7，82.8，79.0。MS MSD trap: m/z 791.0 (M+1) ⁺(accurate mass 790.77).UV-VIS λ max=554nm is in the methyl alcohol.

Embodiment 8-preparation 2 ', 4,5,5 ', 6,7-chlordene-3 ', 6 '-dihydroxy-4 ', 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone

(Fig. 1 g)

Step 1: according to the program that embodiment 1 step 1 is described, the processing tetrachlorophthalic tetrachlorophthalic anhydrid (5g, 17.48mmol), 4-chloro resorcinol (2.78g; 19.23mmol) and the 2-chloro resorcinol (2.78g 19.23mmol), provides the coarse products of 6.2g (to survey about 54% pure 2 ', 4 through HPLC; 5,5 ', 6,7-chlordene-3 '; 6 '-dihydroxy-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone), be used for subsequent step with its collection and as mixture.

Step 2: according to the program that embodiment 1 step 2 is described, iodate 2 ', 4,5; 5 ', 6,7-pentachloro--3 ', 6 '-dihydroxy-3H-spiral shell [isobenzofuran-1; 9 '-xanthene]-3-ketone (6.2g, the compound of 54.4% hope, 6.25mmol); Produce 7.16g (compound of 60.8% hope, 5.50mmol, 88% yield).Via SFC purifying (80g/min total flow, 40% cosolvent (50/50IPA/EtOH0.5%TFA/CO ₂, 140 crust, 254nm is on 3x25cm 5m RegisPack post) separation coarse products (3.37g, the compound of 60.8% hope; 2.59mmol), 2 of 427mg (0.54mmol, the 20.8%SFC rate of recovery) ' is provided, 4; 5,5 ', 6,7-chlordene-3 '; 6 '-dihydroxy-4 ', 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone, be the lightpink solid. ¹H?NMR(300MHz；DMSO?d ₆)δ11.05(s，1H)，10.85(s，1H)，7.60(s，1H)，7.40(s，1H)。 ¹³C?NMR(300MHz；DMSO?d ₆)δ163.1，155.4，154.7，149.6，148.0，146.5，138.7，135.3，134.5，131.3，127.9，126.8，124.7，116.7，110.3，109.0，107.5，82.4，79.6，78.5。MS MSD trap: m/z 790.7 (M+1) ⁺(accurate mass 790.7).UV-VIS λ max=552nm is in the methyl alcohol.

Embodiment 9-is 4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 '; 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone is degraded to 4,5,6; 7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone

(Fig. 1 a and 1i)

To 4,5,6 of 6.5g (6.67mmol), 7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone adds 130mL acetone, 30mL water and 6.6g sodium iodide (44.0mmol).Mixture is heated to reflux continues 82.5 hours, HPLC analyzes and points out that 35.4% title compound is present in the mixture with raw material.Reactant mixture with 130mL water and the dilution of 150mL ethyl acetate, is at room temperature placed and spent the night.Remove organic layer, water layer is used the 100mL ethyl acetate extraction.Organic layer through merging is used brine wash, and is dry on sodium sulphate, produces the 6.73g coarse products, surveys through HPLC and contains 35.4% title compound.Title compound via silica gel chromatography with 66%: 14%: 18%: 4% toluene ∶ diox: hexane: acetate separates from the part of above-mentioned substance.After the continuous chromatography step, 4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone is able to separate, and is red solid (90mg). ¹H?NMR(300MHz；DMSO?d ₆)δ11.26，(s，1H)，10.19(s，1H)，7.56(s，1H)，7.51(s，1H)，6.84(s，1H)。 ¹³C?NMR(300MHz；DMSO?d ₆)δ163.4，159.0，158.3，151.8，150.8，147.2，138.6，136.9，135.1，126.7，124.5，110.6，108.8，101.7，81.2，88.7，77.1。MS MSD trap: m/z 848.8 (M+1) ⁺(accurate mass 847.7).UV-VIS λ max=539nm is in the methyl alcohol.

Embodiment 10-is through incomplete iodate preparation 4,5,6,7-tetrachloro-2 ', 4 ', 5 '-triiodo fluorescein (Fig. 1 h) and allied compound

Add 10g (21.3mmol) tetrachlorofluorescein, 13mL 5MNaOH, 300mL water, 7.08g NaI (47.2mmol) and 16.24g iodine (64mmol) to the 500mL round-bottomed flask.Solution is heated to 50 ℃ continues 6.5 hours, subsequently it is cooled to room temperature.Add sodium hydrogensulfite, adding acetate subsequently is 3.7 until pH.Mixture is used ethyl acetate extraction, and water is brine wash then, and is dry on sodium sulphate, filters and vacuum concentration.18.2g reddish orange foam shape thing be able to separate, be 46%4,5,6,7-tetrachloro-4 ', 5 '-diiodofluorescein and 43% title compound 4,5,6,7-tetrachloro-2 ', the mixture of 4 ', 5 '-triiodo fluorescein is AUC HPLC purity.LCMS MS scans 800-1000:m/z 846.52 (M-1) ^-(accurate mass 847.7).UV-VIS λ max=540nm is among the PBS.Compare with embodiment 1, wherein the iodine of 6 equivalents is converted into the tetraiodide product in 90 ℃ of basal rations, and this reaction uses the heating of milder and the iodine of 3 equivalents to produce the mixture that hangs down the impurity of iodate.This shows how to change the yield of reaction condition with the product of the low iodate of control.

Embodiment 11-is 4,5,6,7-tetrachloro-3 '; 6 '-dihydroxy-2 ', 4 ', 5 ', 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1; 9 '-xanthene]-3-ketone is degraded to 4,5,6,7-tetrachloro-3 '; 6 '-dihydroxy-2 ', 4 ', 5 '-three iodo-7 '-isopropyl-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone (Fig. 1 a and 1s)

At room temperature, to being filled with ZnCl ₂(0.5mL, 0.50mmol), isopropylmagnesium chloride (0.23mL, 0.45mmol) with two (PPh ₃) ₂PdCl ₂The round-bottomed flask of the THF solution of (catalytic amount) adds 200mg (0.21mmol) 4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 ', the 2mL THF solution of 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone.At room temperature agitating solution is 38 hours.Reaction is with 5mL USP water, 0.5mL acetate and the cancellation of 2 5M sulfuric acid.Organic layer is with USP water (3mL) washing, and is dry on sodium sulphate, vacuum concentration.Isolate the orange red solid of 195mg, it is surveyed through HPLC is 4,5,6 of 2.3% title compound and 42.9%, 7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 ', the mixture of 7 '-tetraiodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone.In order to confirm structure, in following embodiment, carry out the separation of title compound.

Embodiment 12-separates 4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 '; 4 ', 5 '-three iodo-7 '-isopropyl-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone and 2,3,4; 5-tetrachloro-6-(6-hydroxyl-2,4,5, the 7-tetraiodo-3-oxo-3H-xanthene-9-yl) benzoic acid disodium salt

(Fig. 1 s to 1t)

The business level Sodium tetraiodotetrachlorofluorescein salt of 118g (0.11mol) is dissolved in 985mL USP water, and is acidified to pH 1-2, slurry is provided with 400mL 1M HCl.This slurry is used the 1800mL ethyl acetate extraction, separate organic layer.Water layer separates organic layer with extra 100mL ethyl acetate extraction.The organic layer that concentrates the warp merging is until the about 1800mL solvent of residue.Then, slurry is handled with the 500mL heptane, filtered, with 250mL heptane wet cake.In 60 ℃ of drying solids, combined with the 760mL diox under blanket of nitrogen in being covered with the flask of paper tinsel then.The gained slurry is heated to 95 ℃, and kept 70 minutes in this temperature.The cooling slurry filters.Under nitrogen, wet cake is washed drying with 2 volume De dioxs.This sample is dissolved in 45mLTHF (not containing inhibitor), is concentrated into 15-20mL, and load on the alumina column that (the 510g neutral alumina is with 300mL 90 acetonitriles: 10 isopropyl alcohols: 1 acetate: 1H ₂O solution).Above-mentioned post is used the same solution wash-out, merge the level that contains product and divide, concentrating under reduced pressure is with the heptane azeotropic drying.Repeat this program for the second time, subsequently consequent solid separated is dissolved in anhydrous THF and is loaded on the preparation of silica gel plate.To prepare plate and use 66% toluene: 14% diox: 18% heptane: 4% acetate wash-out once.Scrape the top bands of a spectrum from the preparation plate,, filter, wash with 20mL THF with the anhydrous THF digestion of 10mL.Vacuum concentrated filtrate provides 59mg title compound, is orange red residue, 85%AUC HPLC purity. ¹H NMR (300MHz; Acetone d ₆) δ 7.67 (s, 1H), 6.99 (s, 1H), 3.27 (m, 1H), 1.12 (d, J=5.4Hz 3H), 1.06 (d, J=5.1Hz, 3H).MS MSD trap: m/z 888.6 (M-1) ^-(accurate mass 887.65).UV-VIS λ max=552nm is in the methyl alcohol.

Only the property intention provides foregoing description presented for purpose of illustration, does not expect to limit the application's invention.

Require scope protection and that hope is protected letters patent to limit as novel teachings accompanying claims.

Claims

1. the method for preparing formula 4 compounds:

R wherein ₁Be Cl or Br independently, R ₂, R ₃, R ₄And R ₅Be I, R ₆Be H, and R ₁₁And R ₁₂Be the equilibrium ion that H maybe can form pharmaceutically acceptable salt independently, said method comprises:

A) being substantially free of cl anion and being substantially free of in the acid solution of the impurity that produces cl anion, with formula 1 compound,

R wherein ₇, R ₈, R ₉And R ₁₀Be H, forming formula 3a midbody compound,

R wherein ₁Be Cl or Br and R independently ₂, R ₃, R ₄, R ₅And R ₆Be H;

B) be substantially free of cl anion and be substantially free of in the solution of the impurity that produces cl anion,, be substantially free of formula 4 compounds of the trans halogenation impurity of formula 4 compounds with formation, wherein R formula 3a midbody compound and iodine combination at least about 4 equivalents ₁Be Cl or Br independently; R ₂, R ₃, R ₄And R ₅In at least one is Cl and remaining is I; R ₆Be H; And R ₁₁And R ₁₂Be the equilibrium ion that H, Na, K, Li maybe can form pharmaceutically acceptable salt independently.

2. the process of claim 1 wherein that the trans halogenation impurity of formula 4 compounds accounts for less than about 0.15 percentage by weight.

3. the process of claim 1 wherein that said trans halogenation impurity comprises is selected from following at least a compound:

2 ', 4,5,6,7-pentachloro--3 ', 6 '-dihydroxy-4 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone; 4,4 ', 5,6,7-pentachloro--3 ', 6 '-dihydroxy-2 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone; 2 ', 4,5,6,7,7 '-chlordene-3 ', 6 '-dihydroxy-4 ', 5 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone; 4,4 ', 5,5 ', 6,7-chlordene-3 ', 6 '-dihydroxy-2 ', 7 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone; 2 ', 4,5,5 ', 6,7-chlordene-3 ', 6 '-dihydroxy-4 ', 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

Their isomery quinoid; And composition thereof.

4. the process of claim 1 wherein that said acid solution comprises the chloride less than about 1500ppm.

5. the method for claim 1; Wherein said acid solution comprise be selected from fusing point less than about 250 ℃ alkyl sulfonic acid or aryl sulfonic acid, fusing point less than about 250 ℃ alkyl carboxylic acid or aryl carboxylic acid, achloride Bronsted acid, the lewis acidic at least a acid of achloride; The prepared product of its polymer-bound, its salt, its aqueous solution; And composition thereof, their lists are used or are made up with methanesulfonic acid.

6. the method for claim 1; Wherein said acid solution comprises at least a acid that is selected from p-methyl benzenesulfonic acid, benzene sulfonic acid, sulfuric acid, TFMS, ethyl sulfonic acid, acetate, propionic acid, trifluoroacetic acid, camphorsulfonic acid; The prepared product of its polymer-bound, its salt, its aqueous solution; And composition thereof, their lists are used or are made up with methanesulfonic acid.

7. the process of claim 1 wherein that said acid solution comprises methanesulfonic acid.

8. the process of claim 1 wherein said formula 2 with formula 1 compound with about 2.5: 1 to about 3.2: 1 ratio combination.

9. the method for claim 1 also is included in the said acid solution and in about 85 ℃ of extremely about 95 ℃ temperature formula 1 and formula 2 compounds is heated about 1 to about 16 hours time period.

10. the method for claim 1 also comprises separate type 3a midbody compound.

11. the method for claim 10, wherein said separation comprise floated 3a midbody compound, collection type 3a midbody compound is with the solvent washing formula 3a midbody compound that is substantially free of cl anion.

12. comprising, the method for claim 11, wherein said solvent be heated to the about 60 ℃ acetone and the mixture of water.

13. the process of claim 1 wherein that the said solution that contains formula 3a midbody compound and iodine comprises less than about 1500ppm chloride.

14. the method for claim 1 also is included in the alkaline solution and will heats about 1 to 24 hour time period through the formula 3a midbody compound and the iodine of combination in about 20 ℃ of extremely about 100 ℃ temperature.

15. the method for claim 14 also is included in the solution and will heats about 1 to 24 hour time period through the formula 3a midbody compound and the iodine of combination in about 70 ℃ of extremely about 95 ℃ temperature.

16. the process of claim 1 wherein that said alkaline solution comprises at least a alkali that is selected from sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium bicarbonate, saleratus and composition thereof.

17. the process of claim 1 wherein that said solution comprises the sodium hydroxide of concentration 0.4 to 1.0M.

18. the method for claim 1 comprises also at least a iodine solubilizer is added the solution that comprises formula 3a midbody compound and iodine that wherein said iodine solubilizer is selected from KI, lithium iodide, sodium iodide and composition thereof.

19. the method for claim 18, wherein said iodine solubilizer comprises about 1 sodium iodide to about 2.5 equivalents.

20. the process of claim 1 wherein that wherein said solution does not contain cl anion, chlorion free radical, hypochlorite, hypochlorous acid or its mixture basically through add oxidant and iodide salt original position generation iodide to solution.

21. the method for claim 1 also is included in the formula of formation 4 compounds and afterwards the iodine trapping agent is added solution, wherein said iodine trapping agent is selected from sodium thiosulfate, potassium thiosulfate, ATS (Ammonium thiosulphate), potassium sulfite, sodium sulphite and composition thereof.

22. the method for claim 21, wherein said iodine trapping agent is a sodium sulphite.

23. the method for claim 22, be added in about 10 ℃ or the lower temperature of wherein said iodine trapping agent are carried out.

24. the process of claim 1 wherein R ₁₁And R ₁₂Comprise sodium.

25. be used for preparing the method for formula 4 compounds

R wherein ₁Be Cl or Br independently, R ₂, R ₃, R ₄And R ₅Be I, R ₆Be H, and R ₁₁And R ₁₂Be the equilibrium ion that H, Na, K, Li maybe can form pharmaceutically acceptable salt independently, be included in and be substantially free of cl anion and be substantially free of in the solution of the impurity that produces cl anion, with formula 3a compound,

Or its quinoid isomer, wherein R ₁Be Cl or Br and R independently ₂, R ₃, R ₄, R ₅And R ₆Be H, and its Chinese style 3a or its quinoid isomer be substantially free of cl anion impurity and the reagent that produces cl anion,, be substantially free of formula 4 compounds of the trans halogenation impurity of formula 4 compounds with formation, wherein R with iodine combination at least about 4 equivalents ₁Be Cl or Br independently, R ₂, R ₃, R ₄And R ₅In at least one is Cl and remaining is I, R ₆Be H, and R ₁₁And R ₁₂Be the equilibrium ion that H, Na, K, Li maybe can form pharmaceutically acceptable salt independently.

26. the method for claim 25, the trans halogenation impurity of its Chinese style 4 compounds accounts for the percentage by weight less than about 0.15.

27. comprising, the method for claim 25, wherein said trans halogenation impurity be selected from following at least a compound:

Its isomery quinoid; And composition thereof.

28. the method for claim 25, wherein said acid solution comprises the chloride less than about 1500ppm.

29. the method for claim 25 also is included in the alkaline solution and will heats about 1 to 24 hour time period through the formula 3a compound and the iodine of combination in about 20 ℃ of extremely about 100 ℃ temperature.

30. the method for claim 29 also is included in the solution and will heats about 2 to 18 hours time period through the formula 3a compound and the iodine of combination in about 70 ℃ of extremely about 95 ℃ temperature.

31. the method for claim 25, wherein said solution comprise at least a alkali that is selected from sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium bicarbonate, saleratus and composition thereof.

32. the method for claim 25, wherein said solution comprise the sodium hydroxide of concentration 0.4 to 1.0M.

33. the method for claim 25 comprises also at least a iodine solubilizer is added the solution that comprises formula 3a compound and iodine that wherein said iodine solubilizer is selected from KI, lithium iodide, sodium iodide and composition thereof.

34. the method for claim 33, wherein said iodine solubilizer comprises about 1 sodium iodide to about 2.5 equivalents.

35. the method for claim 25, wherein through add oxidant and iodide salt original position generation iodide to solution, wherein said solution does not contain cl anion, chlorion free radical, hypochlorite, hypochlorous acid or its mixture basically.

36. the method for claim 25 also is included in the formula of formation 4 compounds and afterwards the iodine trapping agent is added solution, wherein said iodine trapping agent is selected from sodium thiosulfate, potassium thiosulfate, ATS (Ammonium thiosulphate), potassium sulfite, sodium sulphite and composition thereof.

37. the method for claim 36, wherein said iodine trapping agent is a sodium sulphite.

38. the method for claim 37, be added in about 10 ℃ or the lower temperature of wherein said iodine trapping agent are carried out.

39. the method for claim 25, wherein R ₁₁And R ₁₂Comprise sodium.

40. be used for preparing the method for formula 3 compounds

R wherein ₁Be Cl or Br independently, R ₂, R ₃, R ₄And R ₅Be I and R ₆Be H, comprise:

A) in acid solution about 20 ℃ to about 250 ℃ temperature with formula 1 compound,

R wherein ₇, R ₈, R ₉And R ₁₀Be H, to form formula 3a compound;

Separate type 3a midbody compound,

B) in being substantially free of cl anion and the solution that is substantially free of the impurity that produces cl anion with the iodine of formula 3a midbody compound and about 4 equivalents in about 20 ℃ of extremely about 100 ℃ temperature combinations to form formula 3 compounds;

D) the iodine trapping agent is added formula 3 compounds;

E) with formula 3 compound acidifyings to less than about 5 pH, be cooled to less than about 10 ℃ temperature; With

F) separate formula 3 compounds that are substantially free of trans halogenation impurity, wherein R ₁Be Cl or Br independently, R ₂, R ₃, R ₄And R ₅In at least one is Cl and remaining is I, and R ₆Be H.

41. the method for claim 40, the trans halogenation impurity of its Chinese style 3 compounds accounts for the percentage by weight less than about 0.15.

42. comprising, the method for claim 40, wherein said trans halogenation impurity be selected from following at least a compound:

Its isomery quinoid; And composition thereof.

43. the method for claim 40, wherein said acid solution comprises the chloride less than about 1500ppm.

44. the method for claim 40; Wherein said acid solution comprise be selected from fusing point less than about 250 ℃ alkyl sulfonic acid or aryl sulfonic acid, fusing point less than about 250 ℃ alkyl carboxylic acid or aryl carboxylic acid, achloride Bronsted acid, the lewis acidic at least a acid of achloride; The prepared product of its polymer-bound, its salt, its aqueous solution; And composition thereof, their lists are used or are made up with methanesulfonic acid.

45. the method for claim 40; Wherein said acid solution comprises at least a acid that is selected from p-methyl benzenesulfonic acid, benzene sulfonic acid, sulfuric acid, TFMS, ethyl sulfonic acid, acetate, propionic acid, trifluoroacetic acid, camphorsulfonic acid; The prepared product of its polymer-bound, its salt, its aqueous solution; And composition thereof, their lists are used or are made up with methanesulfonic acid.

46. the method for claim 40, wherein said acid solution comprises methanesulfonic acid.

47. the method for claim 40, wherein said formula 2 with formula 1 compound with about 2.5: 1 to about 3.2: 1 ratio combination.

48. the method for claim 40 also is included in the said acid solution and in about 85 ℃ of extremely about 95 ℃ temperature formula 1 and formula 2 compounds is heated about 1 to about 16 hours time period.

49. the method for claim 40 also comprises separate type 3a midbody compound.

50. the method for claim 49, wherein said separation comprise floated 3a midbody compound, collection type 3a midbody compound, and with the solvent washing formula 3a midbody compound that is substantially free of cl anion.

51. comprising, the method for claim 50, wherein said solvent be heated to the about 60 ℃ acetone and the mixture of water.

52. the method for claim 40, the solution that wherein contains formula 3a midbody compound comprises the chloride less than about 1500ppm.

53. the method for claim 40 also is included in the alkaline solution and will heats about 1 to 24 hour time period through the formula 3a midbody compound and the iodine of combination in about 20 ℃ of extremely about 100 ℃ temperature.

54. the method for claim 53 also is included in the solution and will heats about 1 to 24 hour time period through the formula 3a midbody compound and the iodine of combination in about 70 ℃ of extremely about 95 ℃ temperature.

55. the method for claim 40, wherein the formula 3a midbody compound of combination and the solution of iodine comprise at least a alkali that is selected from sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium bicarbonate, saleratus and composition thereof.

56. the method for claim 40, wherein said solution comprise the sodium hydroxide of concentration 0.4 to 1.0M.

57. the method for claim 40 comprises also at least a iodine solubilizer adding is comprised through the formula 3a midbody compound of combination and the solution of iodine that wherein said iodine solubilizer is selected from KI, lithium iodide, sodium iodide and composition thereof.

58. the method for claim 57, wherein said iodine solubilizer comprises about 1 sodium iodide to about 2.5 equivalents.

59. the method for claim 40; Wherein through add oxidant and iodide salt original position generation iodide to the solution that contains formula 3a midbody compound, wherein said solution does not contain cl anion, chlorion free radical, hypochlorite, hypochlorous acid or its mixture basically.

60. the method for claim 40 also is included in the formula of formation 3 compounds and afterwards the iodine trapping agent is added solution, wherein said iodine trapping agent is selected from sodium thiosulfate, potassium thiosulfate, ATS (Ammonium thiosulphate), potassium sulfite, sodium sulphite and composition thereof.

61. the method for claim 60, wherein said iodine trapping agent is a sodium sulphite.

62. the method for claim 61, wherein be added in about 10 ℃ or the lower temperature of iodine trapping agent are carried out.

63. the method for claim 40, wherein acidifying comprises the solution that not chloride anionic acid adding is contained formula 3.

64. the method for claim 63, wherein said not chloride anionic acid comprise the sulfuric acid of about 1 percentage of concentration to about 5 percentages, and wherein add said sulfuric acid to be enough to the regulating amount of said pH value of solution to about pH 2 to about pH 5.

65. the method for claim 64, wherein with said pH regulator to about 3.

66. the method for claim 40, wherein said separation comprise formula 3 compounds are suspended in the solvent that is selected from water, dimethyl formamide, acetone and composition thereof; Vacuum filtration; With the consequent filtrating of solvent washing that is selected from water, dimethyl formamide, acetone and composition thereof.

67. the method for claim 40 is wherein incited somebody to action wherein R ₁Be Cl or Br independently, R ₂, R ₃, R ₄, R ₅Be I and R ₆Formula 3 compounds that are H are converted into formula 4 quinoids, wherein R ₁₁And R ₁₂Be the equilibrium ion that H maybe can form pharmaceutically acceptable salt independently.

68. the method for claim 67, wherein R ₁₁And R ₁₂Comprise sodium.

69. formula 4 compounds, wherein R ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄, R ₅At least one substituting group be that I and at least one substituting group are Cl or H; R ₆Be H or C independently ₁-C ₄Alkyl; R ₁₁And R ₁₂Be the equilibrium ion that H maybe can form pharmaceutically acceptable salt independently; And all

Dynamic isomer; Its atropisomer, its closed lactone derivatives, its enantiomer, its pharmaceutically acceptable salt, and composition thereof, be used for the chemotherapeutic treatment that the human and animal organizes.

70. the medicine that is used for using in the body, it comprises formula 4 compounds of treating effective dose

R wherein ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄, R ₅At least one substituting group be that I and at least one substituting group are Cl or H; R ₆Be H or C independently ₁-C ₄Alkyl; R ₁₁And R ₁₂Be the equilibrium ion that H maybe can form pharmaceutically acceptable salt independently; And whole dynamic isomers; Its atropisomer; Its closed lactone derivatives; Its enantiomer; Its pharmaceutically acceptable salt and composition thereof; Wherein said medicine is suitable for use in the chemotherapeutic treatment of human and animal's tissue.

71. the medicine of claim 70, wherein said formula 4 compounds account for about 0.001% to less than about 20% weight.

72. comprising, the medicine of claim 71, wherein said formula 4 compounds be selected from following compound:

4,4 ', 5,6,7-pentachloro--3 ', 6 '-dihydroxy-2 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

2 ', 4,5,6,7-pentachloro--3 ', 6 '-dihydroxy-4 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

4,4 ', 5,5 ', 6,7-chlordene-3 ', 6 '-dihydroxy-2 ', 7 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone;

2 ', 4,5,6,7,7 '-chlordene-3 ', 6 '-dihydroxy-4 ', 5 '-two iodo-3H-spiral shell [isobenzofuran-1,9 '-xanthene]-3-ketone;

2 ', 4,5,5 ', 6,7-chlordene-3 ', 6 '-dihydroxy-4 ', 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

4,5,6, the 7-tetrabromobisphenol '-chloro-3 ', 6 '-dihydroxy-4 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

4,5,6,7-tetrabromo-4 '-chloro-3 ', 6 '-dihydroxy-2 ', 5 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

4,5,6, the 7-tetrabromobisphenol ', 5 '-two chloro-3 ', 6 '-dihydroxy-4 ', 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

4,5,6,7-tetrabromo-4 ', 5 '-two chloro-3 ', 6 '-dihydroxy-2 ', 7 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

4,5,6, the 7-tetrabromobisphenol ', 7 '-two chloro-3 ', 6 '-dihydroxy-4 ', 5 '-two iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

4,5,6,7-tetrabromo-3 ', 6 '-dihydroxy-2 ', 4 ', 5 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

4,5,6,7-tetrabromo-3 ', 6 '-dihydroxy-2 ', 4 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 5 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

4,5,6,7-tetrachloro-3 ', 6 '-dihydroxy-2 ', 4 ', 7 '-three iodo-3H-spiral shells [isobenzofuran-1,9 '-xanthene]-3-ketone;

Its isomer; Its salt; And composition thereof.

73. the medicine of claim 70 also is included in position R ₁₁Or R ₁₂One of or both be coupled at least one targeting moiety of formula 4 compounds, R in formula 4 compounds ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄, R ₅At least one substituting group be that I and at least one substituting group are Cl or H; R ₆Be H or C independently ₁-C ₄Alkyl; And whole dynamic isomers; Its atropisomer; Its closed lactone derivatives; Its enantiomer; Its pharmaceutically acceptable salt and composition thereof.

74. the medicine of claim 73, wherein said targeting moiety are selected from DNA (DNA), ribonucleic acid (RNA), amino acid, albumen, antibody, part, hapten; Carbohydrate receptor, carbohydrate compounding ingredient, lipid acceptor, lipid compounding ingredient, protein acceptor, protein compounding ingredient, chelating agent; Encapsulation vehicle, short chain aliphatic hydrocarbon, long chain aliphatic hydrocarbon, aromatic hydrocarbons, aldehyde, ketone, alcohol; Ester, acid amides, amine, nitrile, azide, hydrophilic segment, hydrophobic part and composition thereof.

75. the medicine of claim 70, wherein said medicine is prepared in the delivery vector that is selected from liquid, semisolid, solid, aerosol and composition thereof.

76. the medicine of claim 75, wherein said delivery vector is selected from aqueous suspension, non-aqueous suspension, solution, creme, ointment, gel, syrup, suppository, tablet, capsule and droplet spray agent.

77. the medicine of claim 70, wherein said delivery vector also comprise the auxiliary agent that is selected from excipient, stabilizing agent, emulsifier, dispersant, preservative, buffer, electrolyte, tissue penetration agent, organizes softening agents and composition thereof.

78. the medicine of claim 70, it is suitable for use in the chemotherapeutic treatment of disease, and said disease is the disease that influences skin and relevant organ; Influence the disease of oral cavity and digestive tract and relevant organ; Influence the disease of urethra and genital tract and relevant organ, influence the disease of respiratory tract and relevant organ, influence the disease of the circulatory system and relevant organ; Influence the disease of head and neck; Influence the disease of endocrine system and lymphoreticular system and relevant organ, influence the disease of connective tissue, influence is in the disease and the disease that involves microorganism, virus, fungi or parsitism of the tissue surface of intra-operative exposure.

79. the medicine of claim 78, wherein said chemotherapeutic treatment comprise the following treatment pattern that is selected from: intravenous injection, intraperitoneal injection, intramuscular injection, intracranial injection, intratumor injection, last intracutaneous injection, dermal delivery; Through the oesophagus administration, administration in the abdomen, administration in the appendix, intra-arterial administration, administration in the intra-articular administration, bronchi, administration in the cheek, administration in the capsule; Administration in the administration in the orifice of the stomach, cartilage, administration in the intracavitary administration, brain, administration in the colon, administration in the intradermal administration, capsule, administration in the corium; Administration in the pipe, intraduodenal administration, administration in the bundle, administration in the fat, interior administration is split in administration in the silk, administration in the intragastric administration, gland; Administration in the administration in the administration in the administration in the administration in the administration in the administration in the administration in the liver, intestines, plate, focus, ligament, tongue, breast, marrow; Administration in the meninx, administration in the cardiac muscle, intranasal administration, eye drops, administration in the art, administration in mouthful, administration in the bone, administration in the ovary; In the administration in the pancreas, wall/and the interior administration in top district, administration in the pelvis, administration in the pericardium, administration in the perineum, administration in the intraperitoneal administration, placenta, the interior administration of pleura; Administration in the administration in the pons, prostate, administration in the feeding drug into pulmones, vertebra, administration in the drop rectum with drug, kidney, administration in the sclera, administration in the scrotum; Administration in the administration in the administration in the administration in the administration in the administration in the administration in the administration in the joint, sella turcica, backbone, spleen, breastbone, matrix, synovial membrane, shank; Administration in the testis, intrathoracic administration, administration in the almond vivo medicine-feeding, tracheae, administration in the pipe, administration in the tympanum, administration in the ureter; Administration in the urethra, intrauterine administration, intravaginal administration, administration in the intravascular administration, the heart/ventricles of the brain, administration, intravesical administration and glass vivo medicine-feeding in the backbone.

80. the purposes of formula 4 compounds in the preparation medicine, said medicine is suitable for use in the chemotherapeutic treatment of human and animal's tissue,

R wherein ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄, R ₅At least one substituting group be that I and at least one substituting group are Cl or H; R ₆Be H or C independently ₁-C ₄Alkyl; And whole dynamic isomers; Its atropisomer; Its closed lactone derivatives; Its enantiomer; Its pharmaceutically acceptable salt; And composition thereof.

81. formula 4 compounds are used for the purposes of chemotherapeutic treatment of the disease of human and animal tissue,

82. the purposes of formula 4 compounds in the preparation medicine,

R wherein ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄, R ₅At least one substituting group be that I and at least one substituting group are Cl or H; R ₆Be H or C independently ₁-C ₄Alkyl; And whole dynamic isomers; Its atropisomer; Its closed lactone derivatives; Its enantiomer; Its pharmaceutically acceptable salt; And composition thereof;

Said medicine is suitable for treating and is selected from following medical conditions: the disease that influences skin and relevant organ; Influence the disease of oral cavity and digestive tract and relevant organ, influence the disease of urethra and genital tract and relevant organ, influence the disease of respiratory tract and relevant organ; Influence the disease of the circulatory system and relevant organ; Influence the disease of head and neck, influence the disease of endocrine system and lymphoreticular system and relevant organ, influence the disease of connective tissue; The disease of the tissue surface that influence exposes at intra-operative and involve the disease of microorganism, virus, fungi or parsitism.

83. formula 4 compounds

R wherein ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄And R ₅At least one substituting group be that I and at least one substituting group are Cl or H; R ₆Be H or C independently ₁-C ₄Alkyl; And whole dynamic isomers; Its atropisomer; Its closed lactone derivatives; Its enantiomer; Its pharmaceutically acceptable salt; And composition thereof;

Be used to treat the purposes of medical conditions, said medical conditions is selected from the disease that influences skin and relevant organ, influences the disease of oral cavity and digestive tract and relevant organ; Influence the disease of urethra and genital tract and relevant organ; Influence the disease of respiratory tract and relevant organ, influence the disease of the circulatory system and relevant organ, influence the disease of head and neck; Influence the disease of endocrine system and lymphoreticular system and relevant organ; Influence the disease of connective tissue, the disease of the tissue surface that influence exposes at intra-operative and involve the disease of microorganism, virus, fungi or parsitism.

84. the purposes of claim 81, its Chinese style 4 compounds, R ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄, R ₅At least one substituting group be that I and at least one substituting group are Cl or H; R ₆Be H or C independently ₁-C ₄Alkyl; And whole dynamic isomers; Its atropisomer; Its closed lactone derivatives; Its enantiomer; Its pharmaceutically acceptable salt; And composition thereof, account for about 0.001% to less than about 20% weight.

85. the purposes of claim 81 also comprises in the mankind or animal tissue or it presses close to formula 4 compounds that effective dose is treated at the place, wherein R ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄, R ₅At least one substituting group be that I and at least one substituting group are Cl or H; R ₆Be H or C independently ₁-C ₄Alkyl; And whole dynamic isomers; Its atropisomer; Its closed lactone derivatives; Its enantiomer; Its pharmaceutically acceptable salt; And composition thereof.

86. the purposes of claim 85, wherein said comprising: intravenous injection, intraperitoneal injection, intramuscular injection, intracranial injection, intratumor injection, last intracutaneous injection, dermal delivery through being selected from following administration; Through the oesophagus administration, administration in the abdomen, administration in the appendix, intra-arterial administration, administration in the intra-articular administration, bronchi, administration in the cheek, administration in the capsule; Administration in the administration in the orifice of the stomach, cartilage, administration in the intracavitary administration, brain, administration in the colon, administration in the intradermal administration, capsule, administration in the corium; Administration in the pipe, intraduodenal administration, administration in the bundle, administration in the fat, interior administration is split in administration in the silk, administration in the intragastric administration, gland; Administration in the administration in the administration in the administration in the administration in the administration in the administration in the administration in the liver, intestines, plate, focus, ligament, tongue, breast, marrow; Administration in the meninx, administration in the cardiac muscle, intranasal administration, eye drops, administration in the art, administration in mouthful, administration in the bone, administration in the ovary; In the administration in the pancreas, wall/and the interior administration in top district, administration in the pelvis, administration in the pericardium, administration in the perineum, administration in the intraperitoneal administration, placenta, the interior administration of pleura; Administration in the administration in the pons, prostate, administration in the feeding drug into pulmones, vertebra, administration in the drop rectum with drug, kidney, administration in the sclera, administration in the scrotum; Administration in the administration in the administration in the administration in the administration in the administration in the administration in the administration in the joint, sella turcica, backbone, spleen, breastbone, matrix, synovial membrane, shank; Administration in the testis, intrathoracic administration, administration in the almond vivo medicine-feeding, tracheae, administration in the pipe, administration in the tympanum, administration in the ureter, administration in the urethra; The intrauterine administration, intravaginal administration, administration in the intravascular administration, the heart/ventricles of the brain, administration, intravesical administration and glass vivo medicine-feeding in the backbone.

87. be used for the pharmaceutical composition that is suitable for vivo medicine-feeding of chemotherapeutic treatment, it comprises formula 4 compounds,

88. the pharmaceutical composition of claim 87, wherein said formula 4 compounds, wherein R ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄, R ₅At least one substituting group be that I and at least one substituting group are Cl or H; R ₆Be H or C independently ₁-C ₄Alkyl; And whole dynamic isomers; Its atropisomer; Its closed lactone derivatives; Its enantiomer; Its pharmaceutically acceptable salt; And composition thereof, to exist to concentration less than about 20% weight greater than about 0.001%.

89. the pharmaceutical composition of claim 87 also is included in position R ₁₁Or R ₁₂One of or both be coupled at least one targeting moiety of said formula 4 compounds, R in formula 4 compounds ₁Be F, Cl, Br, I, H or C independently ₁-C ₄Alkyl; R ₂, R ₃, R ₄And R ₅Be Cl, H or I independently, wherein be selected from R ₂, R ₃, R ₄, R ₅At least one substituting group be that I and at least one substituting group are Cl or H; R ₆Be H or C independently ₁-C ₄Alkyl; And whole dynamic isomers; Its atropisomer; Its closed lactone derivatives; Its enantiomer; Its pharmaceutically acceptable salt; And composition thereof.

90. the method for chemosynthesis formula 4 compounds,

R wherein ₁Be Cl or Br independently; R ₂, R ₃, R ₄And R ₅Be I; R ₆Be H; R ₁₁And R ₁₂Be the equilibrium ion that H, Na, K, Li maybe can form pharmaceutically acceptable salt independently,

Be included in and be substantially free of in cl anion and the solution that is substantially free of the impurity that produces cl anion formula 3a compound, its Chinese style 3a compound is gone up not chloride anionic impurity and the impurity that produces cl anion basically,

R wherein ₁Be Cl or Br independently; R ₂, R ₃, R ₄, R ₅And R ₆Be H;

With iodine combination, to form formula 4 compounds, its improvement comprises the trans halide derivative that does not have formula 4 compounds in fact, wherein for the trans halogenation impurity of formula 4 compounds, and R ₁Be Cl or Br independently; R ₂, R ₃, R ₄And R ₅In at least one is Cl and remaining is I; R ₆Be H; R ₁₁And R ₁₂Be the equilibrium ion that H, Na, K, Li maybe can form pharmaceutically acceptable salt independently.

91. the method for claim 90, any single trans halogenation impurity of its Chinese style 4 compounds accounts for the percentage by weight less than about 0.15.

92. comprising, the method for claim 90, wherein said trans halogenation impurity be selected from following at least a compound:

Its isomery quinoid; And composition thereof.

93. the method for claim 90, wherein said solution comprises the chloride less than about 1500ppm.

94. the method for claim 90 also is included in the alkaline solution and will heats about 1 to 24 hour time period through the formula 3a compound and the iodine of combination in about 20 ℃ of extremely about 100 ℃ temperature.

95. the method for claim 94 also is included in the solution and will heats about 2 to 18 hours time period through the formula 3a compound and the iodine of combination in about 70 ℃ of extremely about 95 ℃ temperature.

96. the method for claim 90, wherein said solution comprise at least a alkali that is selected from sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium bicarbonate, saleratus and composition thereof.

97. the method for claim 90, wherein said solution comprise the sodium hydroxide of concentration 0.4 to 1.0M.

98. the method for claim 90 comprises also at least a iodine solubilizer is added the solution that comprises formula 3a compound and iodine that wherein said iodine solubilizer is selected from KI, lithium iodide, sodium iodide and composition thereof.

99. the method for claim 98, wherein said iodine solubilizer comprises about 1 sodium iodide to about 2.5 equivalents.

100. the method for claim 90, wherein through add oxidant and iodide salt original position generation iodide to solution, wherein said solution does not contain cl anion, chlorion free radical, hypochlorite, hypochlorous acid or its mixture basically.

101. the method for claim 90 also is included in the formula of formation 4 compounds and afterwards the iodine trapping agent is added solution, wherein said iodine trapping agent is selected from sodium thiosulfate, potassium thiosulfate, ATS (Ammonium thiosulphate), potassium sulfite, sodium sulphite and composition thereof.

102. the method for claim 101, wherein said iodine trapping agent is a sodium sulphite.

103. the method for claim 101, wherein be added in about 10 ℃ or the lower temperature of iodine trapping agent are carried out.

104. the method for claim 90, wherein R ₁₁And R ₁₂Comprise sodium.